Solid forms of a chemokine receptor antagonist and methods of use thereof

ABSTRACT

The citrate salt of (S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol, which may be used in pharmaceutical applications, is disclosed. The crystalline Citrate Salt, including particular single crystal forms and combinations of the single crystalline forms, are also discussed. Mixtures for forming the crystalline salts are discussed. As well, methods of producing the Citrate Salt, and crystalline forms thereof, and using such Citrate Salt, and crystalline forms thereof, in treating diseases associated with aberrant leukocyte recruitment, activation, or recruitment and activation are also discussed.

PRIORITY INFORMATION

The present application claims priority under 35 U.S.C. §119(e) to U.S.Provisional Application No. 60/637,213, filed Dec. 17, 2004, entitled“Solid Forms of a Chemokine Receptor Antagonist and Methods of UseThereof”, the entire contents of which are hereby incorporated byreference.

FIELD OF THE INVENTION

The large-scale manufacturing of a pharmaceutical composition poses manychallenges to the chemist and chemical engineer. While many of thesechallenges relate to the handling of large quantities of reagents andcontrol of large-scale reactions, the handling of the final productposes special challenges linked to the nature of the final activeproduct itself. Not only must the product be prepared in high yield, bestable, and capable of ready isolation, the product must possessproperties that are suitable for the types of pharmaceuticalpreparations in which they are likely to be ultimately used. Thestability of the active ingredient of the pharmaceutical preparationmust be considered during each step of the manufacturing process,including the synthesis, isolation, bulk storage, pharmaceuticalformulation and long-term formulation. Each of these steps may beimpacted by various environmental conditions of temperature andhumidity.

The pharmaceutically active substance used to prepare the pharmaceuticalcompositions should be as pure as possible and its stability onlong-term storage must be guaranteed under various environmentalconditions. This is absolutely essential to prevent the appearance ofunintended degradation products in pharmaceutical compositions, whichdegradation products may be potentially toxic or result simply inreducing the potency of the composition.

A primary concern for the manufacture of large-scale pharmaceuticalcompounds is that the active substance should have a stable crystallinemorphology to ensure consistent processing parameters and pharmaceuticalquality. If an unstable crystalline form is used, crystal morphology maychange during manufacture and/or storage resulting in quality controlproblems, and formulation irregularities. Such a change may affect thereproducibility of the manufacturing process and thus lead to finalformulations which do not meet the high quality and stringentrequirements imposed on formulations of pharmaceutical compositions. Inthis regard, it should be generally borne in mind that any change to thesolid state of a pharmaceutical composition which can improve itsphysical and chemical stability gives a significant advantage over lessstable forms of the same drug.

The present invention relates to forms of a pharmacologically activecompound having activity as a chemokine receptor antagonist and havinghighly preferred properties for use in certain pharmaceuticalformulations.

BACKGROUND OF THE INVENTION

Chemoattractant cytokines, Chemoattractant cytokines or chemokines are afamily of proinflammatory mediators that are released by a wide varietyof cells to promote recruitment and activation of cells such as T and Blymphocytes, eosinophils, basophils, and neutrophils (Luster et al. NewEng. J. Med, 1998, 338, 436). The chemokines are related in primarystructure and contain four conserved cysteines, which form disulfidebonds. The chemokine family includes the C-X-C chemokines(α-chemokines), and the C-C chemokines (β-chemokines), in which thefirst two conserved cysteines are separated by an intervening residue,or are adjacent, respectively (Baggiolini, M. and Dahinden, C. A.,Immunology Today, 1994, 15, 127).

Chemokines exert their biological activity by binding to specificcell-surface receptors belonging to the family of G-protein-coupledseven-transmembrane-domain proteins (Horuk, Trends Pharm. Sci. 1994, 15,159) which are termed “chemokine receptors”. On binding their cognateligands, chemokine receptors then transduce signals important for thedevelopment and trafficking of specific leukocyte subsets (Baggiolini,et. al., Nature 1994, 15, 365). The chemokines and their cognatereceptors have been implicated as being important mediators ofinflammatory, and allergic diseases, disorders, and conditions, as wellas autoimmune pathologies such as rheumatoid arthritis andatherosclerosis (see, Carter, Current Opinion in Chemical Biology 2002,6, 510; Trivedi et al., Ann. Reports Med. Chem. 2000, 35, 191; Saunderset al., Drug Disc. Today 1999, 4, 80; and Premack et al., NatureMedicine, 1996, 2, 1174). Chemokines and their cognate receptors havealso been implicated in the development of cancer and osteolytic bonedisorders (see, Leukemia 2003, 17, 203; J. Bone Miner. Res. 2002, 19,2065; J. Cell. Biochem. 2002, 87, 386; J. Cell. Physiol. 2000, 183, 196;Exp. Hematol. 2005, 33, 272; J. Clin. Invest. 2001, 108, 1833; Cancer2003, 97, 813; Blood 2003, 102, 311).

Accordingly, agents that block the interaction of chemokines with theircognate receptors are useful in treating inflammatory, allergic, andautoimmune diseases, disorders, or conditions, and are also useful inthe treatment of cancer and osteolytic bone disorders caused by aberrantactivation of leukocytes or lymphocytes.

U.S. patent application No. US2002/0169155 and International PublicationNumber WO 03/045942, both entitled “Chemokine Receptor Antagonists andMethods of Use Thereof”, disclose compounds that exhibit an inhibitoryeffect on the chemokine receptor CCR1. These applications additionallydisclose methods for the preparation of these compounds, pharmaceuticalcompositions containing these compounds, and methods for the prophylaxisand therapy of diseases, disorders, or conditions associated withaberrant leukocyte recruitment and/or activation, including but notlimited to rheumatoid arthritis and multiple sclerosis.

(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol(II) is also specifically disclosed:

The structure and synthesis of the free-base amorphous form of thiscompound is provided in the working examples in US2002/0169155 and WO03/045942, and only a general discussion of a wide variety of salts isdisclosed. These applications do not disclose specific salts orcrystalline forms of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol(II).

When a compound crystallizes from a solution or slurry, it maycrystallize with different spatial lattice arrangements, a propertyreferred to as “polymorphism.” Each of the crystal forms is a“polymorph.” While polymorphs of a given substance have the samechemical composition, they may differ from each other with respect toone or more physical properties, such as solubility and dissociation,true density, melting point, crystal shape, compaction behavior, flowproperties, and/or solid state stability.

As described generally above, the polymorphic behavior of drugs can beof crucial importance in pharmacy and pharmacology. The differences inphysical properties exhibited by polymorphs affect practical parameterssuch as storage stability, compressibility and density (important informulation and product manufacturing), and dissolution rates (animportant factor in determining bio-availability). Differences instability can result from changes in chemical reactivity (e.g.,differential oxidation, such that a dosage form discolors more rapidlywhen it is one polymorph than when it is another polymorph) ormechanical changes (e.g., tablets crumble on storage as a kineticallyfavored polymorph converts to thermodynamically more stable polymorph)or both (e.g., tablets of one polymorph are more susceptible tobreakdown at high humidity). In addition, the physical properties of thecrystal may be important in processing: for example, one polymorph mightbe more likely to form solvates that cause the solid form to aggregateand increase the difficulty of solid handling, or might be difficult tofilter and wash free of impurities (i.e., particle shape and sizedistribution might be different between one polymorph relative toother).

While drug formulations having improved chemical and physical propertiesare desired, there is no predictable means for preparing new drug forms(e.g., polymorphs) of existing molecules for such formulations. Thesenew forms would provide consistency in physical properties over a rangeof environments common to manufacturing and composition usage. In theinstant case, no art describes a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,and crystalline forms thereof. More particularly, no art describes acitrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,and crystalline forms thereof, that have unexpected properties that areuseful for large-scale manufacturing, pharmaceutical formulation, andstorage.

SUMMARY OF THE INVENTION

The present invention is directed to the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,and crystalline forms thereof. Those forms also have unexpectedproperties that are useful for large-scale manufacturing, pharmaceuticalformulation, and storage. The present invention also providespharmaceutical compositions comprising said salt, and crystalline formsthereof; methods for the preparation of said citrate salt andcrystalline forms thereof; and methods for uses of these salts andcrystalline forms thereof for the treatment of a variety of diseases,disorders or conditions as described herein.

The present invention shall be more fully discussed with the aid of thefollowing figures and detailed description below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a X-ray powder diffraction pattern from a measurement ona sample of Form A using CuKα₁ radiation, consistent with an embodimentof the invention.

FIG. 2 depicts isothermal X-ray powder diffraction patterns frommeasurements on a sample of Form A using CoKα₁ (λ=1.7890 Å) and CoKα₂(λ=1.7929 Å) radiation taken at temperatures from −80° C. to 190° C. atintervals of 10° C., in accord with an embodiment of the invention.

FIG. 3 depicts a thermal gravimetric analysis result from a measurementof a sample of Form A taken at a heating rate of 5° C./min in accordwith an embodiment of the invention.

FIG. 4 depicts a differential scanning calorimetry result from ameasurement of a sample of Form A taken at a heating rate of 5° C./minin accord with an embodiment of the invention.

FIG. 5 depicts a water sorption/desorption isotherm result from ameasurement of a sample of Form A taken at 25° C. in accord with anembodiment of the invention.

FIG. 6 depicts a X-ray powder diffraction pattern from a measurement ona sample of Form B using CuKα₁ radiation, consistent with an embodimentof the invention.

FIG. 7 depicts isothermal X-ray powder diffraction patterns frommeasurements on a sample of Form B using CoKα₁ (λ=1.7890 Å) and CoKα₂(λ=1.7929 Å) radiation taken at temperatures from −80° C. to 190° C. atintervals of 10° C., in accord with an embodiment of the invention.

FIG. 8 depicts a thermal gravimetric analysis result from a measurementof a sample of Form B taken at a heating rate of 5° C./min in accordwith an embodiment of the invention.

FIG. 9 depicts a differential scanning calorimetry result from ameasurement of a sample of Form A taken at a heating rate of 5° C./minin accord with an embodiment of the invention.

FIG. 10 depicts a water sorption/desorption isotherm result from ameasurement of a sample of Form B taken at 25° C. in accord with anembodiment of the invention.

FIG. 11 depicts X-ray powder diffraction patterns from measurementsusing CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiation on a sample ofForm A and two samples containing Forms A and C, consistent with anembodiment of the invention.

FIG. 12 depicts isothermal X-ray powder diffraction patterns frommeasurements using CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiationon a sample containing Forms A and C taken at temperatures from ambientto 210° C. at intervals of 10° C., in accord with an embodiment of theinvention.

FIG. 13 depicts a thermal gravimetric analysis result from a measurementof a sample containing Forms A and C taken at a heating rate of 5°C./min in accord with an embodiment of the invention.

FIG. 14 depicts a differential scanning calorimetry result from ameasurement of a sample containing Forms A and C taken at a heating rateof 5° C./min in accord with an embodiment of the invention.

FIG. 15 depicts a water sorption/desorption isotherm result from ameasurement of a sample containing Forms A and C taken at 25° C. inaccord with an embodiment of the invention.

FIG. 16 depicts a X-ray powder diffraction patterns from measurementsusing CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiation on a sample ofForm A, a sample containing Forms A and C, and a sample containing FormsA and D, consistent with an embodiment of the invention.

FIG. 17 depicts isothermal X-ray powder diffraction patterns frommeasurements using CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiationon a sample containing Forms A and D taken at temperatures from ambientto 210° C. at intervals of 10° C., in accord with an embodiment of theinvention.

FIG. 18 depicts a thermal gravimetric analysis result from a measurementof a sample containing Forms A and D taken at a heating rate of 5°C./min in accord with an embodiment of the invention.

FIG. 19 depicts a differential scanning calorimetry result from ameasurement of a sample containing Forms A and D taken at a heating rateof 0.3° C./min in accord with an embodiment of the invention.

FIG. 20 depicts a water sorption/desorption isotherm result from ameasurement of a sample containing Forms A and D taken at 25° C. inaccord with an embodiment of the invention.

FIG. 21 depicts a X-ray powder diffraction pattern from a measurement ona sample of Form E using CuKα₁ radiation, consistent with an embodimentof the invention.

FIG. 22 depicts isothermal X-ray powder diffraction patterns frommeasurements on a sample of Form E using CoKα₁ (λ=1.7890 Å) and CoKα₂(λ=1.7929 Å) radiation taken at temperatures from ambient to 210° C. atintervals of 10° C., in accord with an embodiment of the invention.

FIG. 23 depicts a thermal gravimetric analysis result from a measurementof a sample of Form E taken at a heating rate of 5° C./min in accordwith an embodiment of the invention.

FIG. 24 depicts a differential scanning calorimetry result from ameasurement of a sample of Form E taken at a heating rate of 5° C./minin accord with an embodiment of the invention.

FIG. 25 depicts a water sorption/desorption isotherm result from ameasurement of a sample of Form E taken at 25° C. in accord with anembodiment of the invention.

FIG. 26 depicts a X-ray powder diffraction patterns from measurementsusing CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiation on a sample ofForm E and a sample containing Forms E, F, and G, consistent with anembodiment of the invention.

FIG. 27 depicts isothermal X-ray powder diffraction patterns frommeasurements using CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiationon a sample containing Forms E, F, and G taken at temperatures fromambient to 210° C. at intervals of 10° C., in accord with an embodimentof the invention.

FIG. 28 depicts a thermal gravimetric analysis result from a measurementof a sample containing Forms E, F, and G taken at a heating rate of 5°C./min in accord with an embodiment of the invention.

FIG. 29 depicts a differential scanning calorimetry result from ameasurement of a sample containing Forms E, F, and G taken at a heatingrate of 5° C./min in accord with an embodiment of the invention.

FIG. 30 depicts a water sorption/desorption isotherm result from ameasurement of a sample containing Forms E, F, and G taken at 25° C. inaccord with an embodiment of the invention.

DETAILED DESCRIPTION OF THE INVENTION

As used above, and throughout the description of the invention, thefollowing terms, unless otherwise indicated, shall be understood to havethe following meanings.

“Citrate Salt” is meant to describe the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,and has the structure of formula III.

As used herein, “crystalline” refers to a solid having a highly regularchemical structure. In particular, a crystalline Citrate Salt may beproduced as one or more single crystalline forms of the Citrate Salt.For the purposes of this application, the terms “single crystallineform” and “polymorph” are synonymous; the terms distinguish betweencrystals that have different properties (e.g., different XRPD patterns,different DSC scan results). Pseudopolymorphs are typically differentsolvates of a material, and thus their properties differ from oneanother. Thus, each distinct polymorph and pseudopolymorph of theCitrate Salt is considered to be a distinct single crystalline formherein.

“Substantially crystalline” refers to Citrate salts that may be at leasta particular weight percent crystalline. Particular weight percentagesare 10%, 20%, 30% 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%. In some embodiments, substantiallycrystalline refers to Citrate Salts that are at least 70% crystalline.In other embodiments, substantially crystalline refers to Citrate Saltsthat are at least 90% crystalline.

“Form A” is meant to describe a crystalline form of a compound offormula III that may be characterized using distinguishing data.Exemplary data is found in FIGS. 1, 2, 3, 4, and 5, and in Tables 1 and2.

“Form B” is meant to describe a crystalline form of a compound offormula III that may be characterized using distinguishing data.Exemplary data is found in FIGS. 6, 7, 8, 12, 13, and in Tables 3 and 4.

The term “solvate or solvated” means a physical association of acompound of this invention with one or more solvent molecules. Thisphysical association includes hydrogen bonding. In certain instances thesolvate will be capable of isolation, for example when one or moresolvent molecules are incorporated in the crystal lattice of thecrystalline solid. “Solvate or solvated” encompasses both solution-phaseand isolable solvates. Representative solvates include, for example, ahydrate, ethanolates or a methanolate.

The term “hydrate” is a solvate wherein the solvent molecule is H₂O thatis present in a defined stoichiometric amount, and may for example,include hemihydrate, monohydrate, dihydrate, or trihydrate.

The term “mixture” is used to refer to the combined elements of themixture regardless of the phase-state of the combination (e.g., liquidor liquid/crystalline).

The term “seeding” is used to refer to the addition of a crystallinematerial to initiate recrystallization.

A “subject” is preferably a bird or mammal, such as a human, but canalso be an animal in need of veterinary treatment, e.g., domesticanimals (e.g., dogs, cats, and the like), farm animals (e.g., cows,sheep, fowl, pigs, horses, and the like) and laboratory animals (e.g.,rats, mice, guinea pigs, and the like).

An “effective amount” of the Citrate Salt is an amount which results inthe inhibition of one or more processes mediated by the binding of achemokine to a receptor in a subject with a disease associated withaberrant leukocyte recruitment and/or activation. Examples of suchprocesses include leukocyte migration, integrin activation, transientincreases in the concentration of intracellular free calcium [Ca²⁺]_(i)and granule release of proinflammatory mediators. Alternatively, an“effective amount” of the Citrate Salt is a quantity sufficient toachieve a desired therapeutic and/or prophylactic effect, such as anamount which results in the prevention of or a decrease in the symptomsassociated with a disease associated with aberrant leukocyte recruitmentand/or activation.

As used herein, “pro-inflammatory cells” includes but is not limited toleukocytes, since chemokine receptors can be expressed on other celltypes, such as neurons and epithelial cells.

In one aspect, the present invention is directed to a citrate salt ofthe compound(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.Accordingly, the present invention provides a compound having structuralformula (III):

Provided herein is an assortment of characterizing information todescribe the citrate salt forms of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.It should be understood, however, that not all such information isrequired for one skilled in the art to determine that such particularform is present in a given composition, but that the determination of aparticular form can be achieved using any portion of the characterizinginformation that one skilled in the art would recognize as sufficientfor establishing the presence of a particular form, e.g., even a singledistinguishing peak can be sufficient for one skilled in the art toappreciate that such particular form is present.

In some embodiments, the Citrate Salt is substantially crystalline.Non-limiting examples of crystalline Citrate Salts include a singlecrystalline form of the Citrate Salt (e.g., Form A); a mixture ofdifferent single crystalline forms (e.g., a mixture of Form A and B, amixture of any combination of Forms A, C, and D, a mixture of anycombination of Forms B, E, F, G, and H); and a mixture of one or moresingle crystalline forms that excludes one or more designated singlecrystalline forms (e.g., a mixture of crystalline forms of the CitrateSalt excluding Form A). An embodiment of the invention is also directedto a Citrate Salt that excludes one or more designated singlecrystalline forms from a particular weight percentage of the CitrateSalt (e.g., the Citrate Salt being at least 90% by weight other thanForm A). Particular weight percentages may be 10%, 20%, 30% 40%, 50%,60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, 99.5%, 99.9%, or any percentage between 10% and100%.

Alternatively, embodiments of the invention are directed to acrystalline Citrate Salt, wherein at least a particular percentage byweight of the crystalline Citrate Salt is a specific single crystallineform, a combination of particular crystalline forms, or excludes one ormore particular crystalline forms. Particular weight percentages may be10%, 20%, 30% 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%.

Other embodiments of the invention are directed to the Citrate Saltbeing a single crystalline form, or being substantially a designatedsingle crystalline form. The single crystalline form may be a particularpercentages by weight of the Citrate Salt. Particular weight percentagesare 10%, 20%, 30% 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%. When a particular percentage by weightof a citrate salt is a single crystalline form, the remainder of thecitrate salt is some combination of amorphous form of the Citrate Salt,and one or more crystalline forms of the Citrate Salt excluding thesingle crystalline form.

Examples of a single crystalline form include Form A, B, C, D, E, F, G,and H of the Citrate Salt, as well as descriptions of a singlecrystalline form characterized by one or more properties as discussedherein. The descriptions characterizing the single crystalline forms mayalso be used to describe the mixture of different forms that may bepresent in a crystalline Citrate Salt.

In the following description of particular polymorphs of the citratesalt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,embodiments of the invention may be described with reference to aparticular crystalline “Form” of the Citrate Salt (e.g., Form B).However, the particular crystalline forms of the Citrate Salt may alsobe characterized by one or more of the characteristics of the polymorphas described herein, with or without regard to referencing a particular“Form”.

Form A

Particular embodiments of the invention are directed toward a singlecrystalline form of the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olhaving Form A. In one particular embodiment of the invention, the singlecrystalline form of the Citrate Salt is characterized by a X-ray powderdiffraction (XRPD) pattern substantially similar to FIG. 1, the patterngenerated using CuKα₁ radiation. The single crystalline form of theCitrate Salt may alternatively be characterized by the lines derivedfrom profile fitting the pattern of FIG. 1 as listed in Table 1.

TABLE 1 Indexing of XRPD Pattern of FIG. 1 2-theta multiplicity d-λ_(Cu) K{circumflex over (α)} h k l J spacing (Å) 1.54184 Å 0 1 0 215.1877 5.8191 1 0 0 2 14.0302 6.2996 1 1 0 2 12.3910 7.1340 −1 1 0 29.0102 9.8165 0 0 1 2 8.4624 10.4538 0 −1 1 2 8.2004 10.7887 1 2 0 27.7576 11.4064 0 2 0 2 7.5939 11.6533 −1 −1 1 2 7.4446 11.8878 1 0 1 27.4434 11.8897 2 1 0 2 7.2834 12.1519 −1 0 1 2 7.0641 12.5305 2 0 0 27.0151 12.6184 0 1 1 2 6.7840 13.0501 1 −1 1 2 6.7639 13.0892 1 1 1 26.6066 13.4022 0 −2 1 2 6.3956 13.8465 −1 −2 1 2 6.3605 13.9232 2 2 0 26.1955 14.2959 −1 2 0 2 5.9522 14.8835 −2 1 0 2 5.7297 15.4649 −1 1 1 25.7068 15.5275 −2 −1 1 2 5.6454 15.6972 2 0 1 2 5.5646 15.9267 2 1 1 25.4031 16.4060 1 −2 1 2 5.3966 16.4261 −2 −2 1 2 5.3339 16.6204 1 3 0 25.3156 16.6778 −2 0 1 2 5.2505 16.8863 1 2 1 2 5.2383 16.9259 0 2 1 25.1189 17.3236 2 −1 1 2 5.0633 17.5153 0 3 0 2 5.0626 17.5180 −1 −3 1 24.9960 17.7531 3 1 0 2 4.9178 18.0380 2 3 0 2 4.8839 18.1641 0 −3 1 24.8396 18.3318 2 2 1 2 4.7201 18.8000 3 2 0 2 4.6805 18.9608 3 0 0 24.6767 18.9760 −2 −3 1 2 4.5839 19.3641 −2 2 0 2 4.5051 19.7060 −2 1 1 24.4792 19.8213 −1 2 1 2 4.4702 19.8613 −1 3 0 2 4.3518 20.4077 0 −1 2 24.3301 20.5110 2 −2 1 2 4.2880 20.7147 −3 −1 1 2 4.2691 20.8073 1 −3 1 24.2453 20.9254 −3 −2 1 2 4.2356 20.9735 3 1 1 2 4.2350 20.9768 0 0 2 24.2312 20.9958 3 0 1 2 4.1996 21.1555 −1 −1 2 2 4.1702 21.3064 3 3 0 24.1303 21.5144 1 3 1 2 4.1293 21.5197 −3 1 0 2 4.1252 21.5414 1 0 2 24.1188 21.5755 1 −1 2 2 4.1056 21.6458 0 −2 2 2 4.1002 21.6746 −1 −2 2 24.0573 21.9067 −3 0 1 2 3.9946 22.2548 1 4 0 2 3.9893 22.2845 −1 0 2 23.9864 22.3010 0 3 1 2 3.9757 22.3621 3 2 1 2 3.9689 22.4009 −1 −4 1 23.9504 22.5069 2 3 1 2 3.9472 22.5256 −3 −3 1 2 3.9139 22.7197 3 −1 1 23.8833 22.9011 2 4 0 2 3.8788 22.9279 0 1 2 2 3.8619 23.0297 1 1 2 23.8565 23.0623 1 −2 2 2 3.8242 23.2597 −2 −4 1 2 3.8151 23.3162 0 4 0 23.7969 23.4293 0 −4 1 2 3.7901 23.4720 −2 −1 2 2 3.7312 23.8482 −2 2 1 23.7247 23.8906 −2 −2 2 2 3.7223 23.9060 2 0 2 2 3.7217 23.9099 −1 −3 2 23.7097 23.9884 4 1 0 2 3.6759 24.2123 0 −3 2 2 3.6686 24.2611 4 2 0 23.6417 24.4431 2 −1 2 2 3.6399 24.4556 −2 3 0 2 3.6096 24.6640 −1 1 2 23.6032 24.7081 2 1 2 2 3.5901 24.7997 −1 3 1 2 3.5794 24.8750 2 −3 1 23.5705 24.9380 −3 1 1 2 3.5547 25.0511 3 3 1 2 3.5381 25.1702 −2 0 2 23.5321 25.2141 3 4 0 2 3.5299 25.2301 −3 2 0 2 3.5251 25.2649 −2 −3 2 23.5096 25.3784 4 0 0 2 3.5076 25.3932 −3 −4 1 2 3.4608 25.7420 1 2 2 23.4463 25.8526 3 −2 1 2 3.4381 25.9148 1 −4 1 2 3.4238 26.0253 4 3 0 23.4205 26.0507 −1 4 0 2 3.4089 26.1408 1 −3 2 2 3.4080 26.1478 −4 −2 1 23.4003 26.2083 0 2 2 2 3.3920 26.2734 4 1 1 2 3.3822 26.3513 2 −2 2 23.3819 26.3532 −4 −1 1 2 3.3610 26.5200 1 4 1 2 3.3423 26.6716 4 0 1 23.3101 26.9353 2 2 2 2 3.3033 26.9922 2 4 1 2 3.2941 27.0688 4 2 1 23.2925 27.0823 −4 −3 1 2 3.2769 27.2141 −1 −4 2 2 3.2747 27.2324 −3 −2 22 3.2658 27.3079 3 0 2 2 3.2341 27.5810 −3 −1 2 2 3.2221 27.6853 −1 −5 12 3.2147 27.7508 0 4 1 2 3.2101 27.7914 −2 1 2 2 3.2083 27.8076 −4 1 0 23.2071 27.8180 0 −4 2 2 3.1978 27.9004 3 1 2 2 3.1929 27.9444 −2 −5 1 23.1864 28.0019 −2 −4 2 2 3.1803 28.0575 1 5 0 2 3.1790 28.0687 −4 0 1 23.1746 28.1085 2 5 0 2 3.1674 28.1740 −1 2 2 2 3.1656 28.1896 −3 −3 2 23.1643 28.2017 3 −1 2 2 3.1343 28.4773 −2 3 1 2 3.1160 28.6482 4 −1 1 23.1041 28.7601 4 4 0 2 3.0978 28.8205 −3 2 1 2 3.0967 28.8309 3 4 1 23.0868 28.9253 0 −5 1 2 3.0831 28.9605 4 3 1 2 3.0752 29.0364 −3 0 2 23.0493 29.2887 2 −3 2 2 3.0421 29.3594 0 5 0 2 3.0375 29.4047 −4 −4 1 23.0368 29.4125 3 2 2 2 3.0254 29.5249 1 3 2 2 3.0174 29.6049 −3 −5 1 23.0100 29.6798 3 5 0 2 3.0074 29.7065 −3 3 0 2 3.0034 29.7464 2 −4 1 22.9993 29.7881 3 −3 1 2 2.9941 29.8414 1 −4 2 2 2.9816 29.9696 −2 4 0 22.9761 30.0258

The single crystalline form of the Citrate Salt may also becharacterized by one or more of the peaks of a XRPD pattern. Forexample, one embodiment of the invention characterizes a singlecrystalline form of the Citrate Salt using at least one linecorresponding to a major peak of the XRPD pattern of FIG. 1. The majorpeaks are labeled A-M on FIG. 1. The major peaks, as identified by theircorresponding 2θ location from Table 1, are: 5.8, 9.8, 11.7, 12.6, 15.5,15.7, 15.9, 17.3, 17.5, 17.8, 18.2, 19.0, and 19.7. The error in the 2θlocations are typically within ±0.1. Other examples of characterizing asingle crystalline form utilize any number of the listed major peaks(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13), or utilizing asubset of the major peaks that do not overlap peaks associated with oneor more other polymorphs (e.g., utilizing one, two, three, four, five,six, seven or more of the major peaks listed above but excluding thepeaks that overlap the peaks of Form B). In an alternative particularembodiment of the invention, the single crystalline form of the CitrateSalt is characterized by one or more of the calculated cell parametersfound in Table 2. The calculated cell parameters are based upon analysisof corresponding XRPD measurements. From the results of the analysis,the crystalline structure is also characterized to be triclinic P1 (Z=1and Z′=2).

In some embodiments, Form A can be characterized by at least one of thex-ray powder diffraction peaks at 2θ angles of 9.8, 11.7, 12.6, 15.5,15.7, 15.9, 17.3, 17.5, 18.2, 19.0, and 19.7.

TABLE 2 Cell Parameters from XRPD Scan of Form A a = 14.754(4) Å b =16.345(0) Å c = 8.676(0) Å α = 102.29(8)° β = 90.56(8)° γ = 72.29(7)° V= 1944.1 Å³ Density = 1.239 (Z = 1 and Z′ = 2)

In another particular embodiment of the invention, the singlecrystalline form of the Citrate Salt is characterized by having astability transition in the range of 150° C. and 160° C. as observedwith controlled temperature X-ray powder diffraction. In FIG. 2, theresults of a series of XRPD scans, measured at temperatures from −80° C.to 190° C. at intervals of 10° C. on a sample of Form A using CoKα₁(λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiation, are presented. It isnoted that from −80° C. to 150° C. the XRPD graphs do not show anappreciable change except for a typical heat-induced dilation as thetemperature changes; the 150° C. measurement is shown in FIG. 2 by thescan 210. Above 150° C., however, the scans at progressively highertemperatures show a continuous evolution, corresponding to a transitionaway from a stable crystalline form between 150° C. and 160° C.

In another embodiment of the invention, Form A is characterized by adecomposition/melting phenomena above 170° C. as observed withcontrolled temperature X-ray powder diffraction. The measurements at180° C. and 190° C. (shown by the scans 220, 230, respectively) in FIG.2 have essentially no structural features, and correspond to adecomposed/melted crystalline Citrate Salt.

Form A is also characterized by a TGA measurement, in other embodimentsof the invention. As shown in FIG. 3, the graph 310 corresponds to theweight of the sample 315 as a function of temperature, while graph 320corresponds to a derived rate of weight loss 325 as a function oftemperature, the measurement being performed at a constant rate oftemperature change of 5° C./min. This polymorph is anhydrous; it has avery small weight loss 311 of about 0.23% (w/w) from ambient temperatureto about 100° C. shown in FIG. 3. The polymorph is also characterized bythe weight loss transition 312 corresponding to a weight loss of about27.5% (w/w) beginning at about 164° C., as interpolated 326 from thederived rate of weight loss curve 320. The inflection point of thisweight loss transition 312, corresponding to the maximum rate of weightloss 327, at 183° C. also characterizes this weight loss transition. Theprecision of all temperatures in all TGA scans herein are within ±3° C.of the stated temperatures.

A DSC measurement of a sample of Form A may be utilized to characterizea single crystalline form of the Citrate Salt in embodiments of theinvention. FIG. 4 depicts the result of a DSC measurement on a sample ofForm A at a heating rate of 5° C./min. The curve 410 depicts the heatflow into the sample as a function of temperature. A particularembodiment of the invention characterizes Form A by the endothermictransition 411 of the curve 410, the transition interpolated 412 tobegin at 169° C., in FIG. 4. The maximum rate of heat loss 413 duringthe transition occurs at 176° C., the temperature being designatedT_(max), also characterizes this transition. The endothermic transitioncorresponds with the melting/decomposition of the crystalline sample.The precision of all temperatures in this DSC scans herein are within±3° C. of the stated temperatures.

A DSC measurement on a sample of Form A performed at a heating rate of1° C./min may also be used to characterize a single crystalline form ofthe Citrate Salt, in accordance with an embodiment of the invention. Insuch a measurement, the DSC scan is characterized by and endothermictransition interpolated to begin at 164° C.; a T_(max)=167° C. alsoserves to characterize the endothermic transition.

Particular embodiments of the invention utilize characterizations ofForm A that refer to results of water sorption/desorption cycling. FIG.5 depicts the result of two consecutive water sorption/desorption cyclesperformed at 25° C. on sample of Form A. The curves depict the percentchange in mass, on a dry crystal basis, of the sample as a function ofrelative humidity.

The overlapping curves indicate the reversibility of the sorptionphenomena. Correspondingly, Form A is also characterized from theobservation that XRPD measurements made before and after cyclingindicate no structural change in the sample.

In one embodiment of the invention, Form A is characterized by a changeof about 1.9% in mass as the relative humidity is changed between 0% and90% relative humidity, as shown in FIG. 5. The % change in mass is knownto within ±0.1%. Alternatively, the embodiment may be characterized by achange of about 0.8 moles of water per mole of anhydrous crystal overthe relative humidity range of 0% to 90%.

Embodiments of the invention also characterize Form A from XRPDmeasurements showing that the crystal does not change form whensubjected to a week long exposure to a relative humidity of 97.5% atambient temperature.

Form B

Form B is a single crystalline form of the Citrate Salt in a compositionconsistent with particular embodiments of the invention. One particularembodiment of the invention is directed towards Form B having asubstantially similar XRPD pattern to what is displayed in FIG. 6. CuKα₁radiation is used to generate the pattern of FIG. 6. Form B isalternatively characterized by the lines derived from profile fittingthe XRPD pattern of FIG. 6, as listed in Table 3.

TABLE 3 Indexing of XRPD Pattern of FIG. 6 2-theta d- λ_(Cu)K{circumflex over (α)} h k l multiplicity J spacing (Å) 1.54184 Å 0 2 02 15.2730 5.7866 1 1 0 4 13.2774 6.6572 1 2 0 4 10.6073 8.3357 1 3 0 48.3781 10.5592 0 1 1 4 8.2924 10.6686 0 4 0 2 7.6365 11.5880 0 2 1 47.5043 11.7929 1 0 1 4 7.4388 11.8970 2 0 0 2 7.3715 12.0061 1 1 1 87.2276 12.2460 2 1 0 4 7.1658 12.3520 1 4 0 4 6.7808 13.0562 1 2 1 86.6877 13.2388 2 2 0 4 6.6387 13.3370 0 3 1 4 6.5772 13.4624 1 3 1 86.0066 14.7481 2 3 0 4 5.9710 14.8365 0 4 1 4 5.7149 15.5053 1 5 0 45.6438 15.7017 2 0 1 4 5.6012 15.8219 2 1 1 8 5.5094 16.0874 1 4 1 85.3286 16.6372 2 4 0 4 5.3036 16.7159 2 2 1 8 5.2587 16.8596 0 6 0 25.0910 17.4194 0 5 1 4 4.9836 17.7979 2 3 1 8 4.9077 18.0755 3 1 0 44.8519 18.2848 1 6 0 4 4.8122 18.4373 1 5 1 8 4.7211 18.7960 2 5 0 44.7038 18.8659 3 2 0 4 4.6781 18.9704 2 4 1 8 4.5165 19.6556 3 3 0 44.4258 20.0628 0 6 1 4 4.3830 20.2606 0 0 2 2 4.3080 20.6173 3 0 1 44.2688 20.8088 0 1 2 4 4.2658 20.8236 3 1 1 8 4.2277 21.0133 1 6 1 84.2013 21.1469 2 6 0 4 4.1891 21.2094 1 7 0 4 4.1843 21.2339 0 2 2 44.1462 21.4311 1 0 2 4 4.1351 21.4895 3 4 0 4 4.1326 21.5028 2 5 1 84.1286 21.5237 3 2 1 8 4.1112 21.6158 1 1 2 8 4.0977 21.6879 1 2 2 83.9914 22.2729 0 3 2 4 3.9675 22.4087 3 3 1 8 3.9368 22.5858 0 7 1 43.8929 22.8438 1 3 2 8 3.8312 23.2169 3 5 0 4 3.8292 23.2292 0 8 0 23.8182 23.2966 2 6 1 8 3.7674 23.6157 1 7 1 8 3.7639 23.6379 2 7 0 43.7551 23.6941 0 4 2 4 3.7521 23.7131 3 4 1 8 3.7261 23.8810 2 0 2 43.7194 23.9248 1 8 0 4 3.6963 24.0766 2 1 2 8 3.6921 24.1041 4 0 0 23.6858 24.1465 4 1 0 4 3.6592 24.3243 1 4 2 8 3.6362 24.4805 2 2 2 83.6138 24.6348 4 2 0 4 3.5829 24.8506 3 6 0 4 3.5358 25.1873 0 5 2 43.5207 25.2969 3 5 1 8 3.4992 25.4550 2 3 2 8 3.4936 25.4962 0 8 1 43.4908 25.5170 4 3 0 4 3.4657 25.7053 2 7 1 8 3.4424 25.8824 1 5 2 83.4244 26.0205 1 8 1 8 3.3969 26.2349 2 8 0 4 3.3904 26.2859 4 0 1 43.3887 26.2995 4 1 1 8 3.3680 26.4637 2 4 2 8 3.3439 26.6586 4 4 0 43.3194 26.8592 4 2 1 8 3.3083 26.9509 1 9 0 4 3.3075 26.9573 0 6 2 43.2886 27.1151 3 6 1 8 3.2710 27.2634 3 7 0 4 3.2630 27.3320 3 0 2 43.2395 27.5341 3 1 2 8 3.2214 27.6916 4 3 1 8 3.2153 27.7454 1 6 2 83.2097 27.7947 2 5 2 8 3.1769 28.0873 3 2 2 8 3.1690 28.1592 0 9 1 43.1578 28.2608 4 5 0 4 3.1559 28.2786 2 8 1 8 3.1549 28.2872 4 4 1 83.0974 28.8236 1 9 1 8 3.0878 28.9156 3 3 2 8 3.0870 28.9230 2 9 0 43.0829 28.9623 0 7 2 4 3.0657 29.1283 0 10 0 2 3.0546 29.2368 3 7 1 83.0515 29.2672 3 8 0 4 3.0151 29.6282 2 6 2 8 3.0033 29.7478 1 7 2 83.0015 29.7657 1 10 0 4 2.9911 29.8720 4 6 0 4 2.9855 29.9293 3 4 2 82.9823 29.9624 4 5 1 8 2.9634 30.1581

Form B is also characterized by one or more of the peaks in a XRPDpattern. For example, one embodiment of the invention characterizes asingle crystalline form of the Citrate Salt using at least one linecorresponding to a major peak of the XRPD pattern of FIG. 6. The majorpeaks are labeled A-M on FIG. 6. The major peaks, as identified by theircorresponding 2θ location in Table 3, are: 5.8, 10.6, 11.6, 12.3, 14.8,15.8, 16.1, 16.7, 17.8, 18.8, 20.6, 21.7, and 24.5. The error in the 2θlocations are typically within ±0.1. Other examples utilize any numberof the listed major peaks (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,or 13), or utilizing a subset of the major peaks that do not overlappeaks associated with one or more other polymorphs (e.g., utilizing one,two, three, four, five, six, seven or more of the major peaks listedabove but excluding the peaks that overlap the peaks of Form A). Inanother alternate embodiment of the invention, Form B is characterizedby one or more of the calculated cell parameters of Table 4, derivedfrom analysis conducted on a XRPD measurement. The crystalline structureis also characterized to be orthorhombic P2₁2₁2₁ (Z=4 and Z′=2).

In some embodiments, Form B can be characterized by at least one of thex-ray powder diffraction peaks at 2θ angles of 10.6, 11.6, 12.3, 14.8,15.8, 16.1, 16.7, 18.8, 20.6, 21.7, and 24.5.

TABLE 4 Cell Parameters from XRPD Scan of Form B a = 14.743(8) Å b =30.546(5) Å c = 8.616(4) Å α = 90° β = 90 γ = 90° V = 3880.6 Å³ Density= 1.241 (Z = 4 and Z′ = 1)

In a particular embodiment of the invention, Form B is characterized bya stability transition between 170° C. and 180° C. as detected fromtemperature controlled XRPD measurements. FIG. 7 depicts XRPDmeasurements on a sample Form B using CoKα₁ (λ=1.7890 Å) and CoKα₂(λ=1.7929 Å) radiation. Each scan represents an isothermal measurementof the sample, the temperatures ranging from −80° C. to 190° C. atintervals of 10° C. Up to 170° C., the XRPD scans at differenttemperatures only display slight variations between each other due totypical heat-induced dilation (e.g., scan 710 corresponds with themeasurement at 170° C.). Scans above 170° C. show alteration of spectralfeatures corresponding to a transition away from a stable crystallineform.

In another embodiment of the invention, Form B is characterized bydecomposition/melting between 180° C. and 190° C. as observed fromisothermal XRPD measurements. This is indicated by the measurement at190° C. (shown by scan 720) being essentially featureless andcorresponding to decomposed/melted Form B.

In a particular embodiment of the invention, Form B is characterized byone or more features of the TGA measurement in FIG. 8. The curve 810graphs the weight of the sample 815 of Form B that is measured as afunction of temperature; corresponding curve 820 graphs the derived rateof weight loss 825 as a function of temperature. The heating rate of themeasurement is 5° C./min. This polymorph is anhydrous. No weight loss isdetected from ambient temperature to 100° C., as shown in FIG. 8. Thesingle crystalline form is also be characterized by the weight losstransition 811 corresponding to a weight loss of about 27.5% (w/w)beginning at about 180° C., as interpolated 826 from the derived rate ofweight loss curve 820. The inflection point of this weight losstransition 811, corresponding to the maximum rate of weight loss 827, at192° C. also characterizes this weight loss transition.

In another particular embodiment of the invention, Form B ischaracterized by one of more features of the DSC measurement depicted inFIG. 12. Using a heating rate of 5° C./min, the curve 1210 graphs theheat flow as a function of temperature. The endothermic transition (orheat flow transition) 1211 of the curve 1210 may be used to characterizethe crystalline form. The transition is characterized by an interpolated1212 beginning temperature at 184° C., and a maximum rate of heat loss1213 during the transition at T_(max)=189° C. The transition correspondswith the melting/decomposition of the Form B sample.

In a related embodiment of the invention, Form B is characterized by aDSC measurement conducted at a heating rate of 1° C./min. In such ameasurement, the DSC scan is characterized by and endothermic transitioninterpolated to begin at 174° C.; a T_(max)=179° C. also serves tocharacterize the endothermic transition.

Other particular embodiments of the invention utilizing Form B maycharacterize the polymorph from a water sorption/desorption cyclingexperiment corresponding to the graph of FIG. 13. FIG. 13 shows theresult of two consecutive water sorption/desorption cycles performed at25° C. on sample of Form B, depicting the percent change in mass, on adry crystal basis, of the sample as a function of relative humidity.

The overlapping curves indicate the reversibility of the sorptionphenomena. Correspondingly, Form B is also be characterized from theobservation that XRPD measurements made before and after cyclingexperiment indicate no structural change in the sample.

In one embodiment of the invention, Form B is characterized by a changeof about 0.9% in mass as the relative humidity is changed between 0% and90% relative humidity, as shown in FIG. 13. The % change in mass isknown to within ±0.1%. Alternatively, the embodiment may becharacterized by a change of about 0.4 moles of water per mole ofanhydrous crystal over the relative humidity range of 0% to 90%.

Embodiments of the invention also characterize Form B from XRPDmeasurements showing that the crystal does not change form whensubjected to a month long exposure to a relative humidity of 97.5% atambient temperature. In addition, Form B is characterized by itsstability when rinsed with water, as observed by XRPD before and afterthe rinsing; this also suggests the single crystalline form ischaracterized as being stable to 100% relative humidity at ambienttemperature.

Form C

In an embodiment of the invention, a single crystalline form of theCitrate Salt is characterized as having Form C. In particular, thesingle crystalline form may be characterized by particular features fromthe XRPD measurements shown in FIG. 11 using CoKα₁ (λ=1.7890 Å) andCoKα₂ (λ=1.7929 Å) radiation. FIG. 11 compares the relative intensity asa function of 2θ for scans 1110, 1120, 1130. Scans 1110 and 1130correspond to measurements of samples including a mixture of Form A andForm C crystals. Scan 1120, corresponding to a sample of pure Form A, isprovided to distinguish features in the XRPD scans 1110, 1130 that areunique to Form C of the Citrate Salt. At least four peaks that appearunique to Form C of the Citrate Salt are designated with a * on scan1110 of FIG. 11. Thus, the single crystalline form of the Citrate Saltis characterized any number of these four peaks.

In another embodiment of the invention, Form C is characterized by aloss of stability between 150° C. and 160° C. as observed withtemperature dependent XRPD measurements. FIG. 12 depicts XRPDmeasurements, using CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å) radiation,on the same sample used to generate scan 1110 in FIG. 11. Each curverepresents an isothermal scan of the sample in a dry nitrogenenvironment; the temperatures ranging from ambient temperature to 210°C. in intervals of 10° C. As the temperature is raised, the peak at2θ˜6.35 begins to lose intensity, the successive scans progressivelyresembling Form A. For the scan 1220 corresponding to T=160° C., thepeaks characteristic of Form C are essentially not present; the samplenow corresponding to crystalline Form A. Further increases intemperature eventually bring about the decomposition/melting of thecrystalline Form A (e.g., scan 1230 corresponding to T=180° C.). Thus,XRPD measurements indicate a conversion of the Form C crystals to Form Abetween 150° C. and 160° C.

Form C may also be characterized by particular features of a TGAmeasurement as shown in FIG. 13. The curve 1310 graphs the weight of thetested sample 1315 including Forms A and C as a function of temperature;corresponding curve 1320 graphs the derived rate of weight loss 1325 asa function of temperature.

In particular, Form C is characterized by the weight loss 1312,interpolated to begin at 157° C. 1321 and having a maximum rate ofweight loss at 162° C. 1322. This weight loss of about 8.22% (w/w) isrelated to the conversion away from Form C. Thus, Form C ischaracterized as a crystal of the Citrate Salt that converts to Form A,beginning at about 157° C. with a maximum rate of weight loss at 162° C.according to TGA. Corresponding measurements made using TGAsimultaneously with DSC, and a TGA measurement coupled with massspectrometry, confirm these results.

A weight loss 1313 corresponds to the decomposition/melting of thesubsequent Form A crystal; the third weight loss of 21.9% (w/w) having amaximum weight loss rate at 183° C. and beginning at 168° C.

The DSC measurement of FIG. 14 also characterize Form C. The curve 1410,generated from testing a sample including Forms A and C at a heatingrate of 5° C./min, shows two strong endothermic transitions. Inparticular, Form C is characterized by the first endothermic transition1411, interpolated to begin at about 161° C. and having a T_(max)=166°C. This is associated with the conversion of crystalline Form C to FormA. The second endothermic transition 1412, interpolated to begin atabout 173° C. and having a T_(max)=178° C., corresponds with themelting/decomposition of Form A.

Alternatively, Form C is be characterized by a DSC measurement on asample at a heating rate of 20° C./min. In particular, the endothermictransition corresponding to a transition between Form C and Form A ischaracterized by an interpolated beginning temperature of 165° C. and aT_(max) of 171° C.

A single crystalline form of the Citrate Salt is also characterized bythe curves of FIG. 15, corresponding to two consecutive watersorption/desorption cycles performed at 25° C. on a sample that includesForms A and C, consistent with an embodiment of the invention. Thegraphs depict the percent change in mass, on a dry crystal basis, of thecrystal as a function of relative humidity. Form C is characterized bybeing stable over a range of relative humidity from 0% to 90% asindicated by the overlap of the curves (i.e., the sorption phenomena isreversible with respect to the sample). XRPD scans of the sample beforeand after the sorption/desorption cycling show no structuralmodification (i.e., the proportion of Form A to Form C crystals isjudged to be constant). Thus, the XRPD scans further characterize thestability of Form C over the relative humidities tested.

Form D

Some embodiments of the invention are directed to a crystalline CitrateSalt including Form D. In particular, Form D is characterized byfeatures of XRPD measurements performed with CoKα₁ (λ=1.7890 Å) andCoKα₂ (λ=1.7929 Å) radiation as shown in FIG. 16. Scans 1610, 1620, 1630graph the relative intensity as a function of 2θ for three samples. Scan1610 corresponds to a sample of Forms A and D. Scans 1620, 1630,corresponding to samples of a combination of Forms A and C and pure FormA, respectively, are used to derive the unique features of scan 1610corresponding with Form D. In one embodiment, Form D is characterized byany number of the peaks designated with a * on scan 1610 of FIG. 16.

Form D is characterized by a loss of stability between 40° C. and 50° C.as observed by temperature controlled XRPD measurements. FIG. 17presents isothermal XRPD measurements on sample containing Forms A and Dat temperatures from ambient to 210° C., in steps of 10° C. in a drynitrogen environment. For scans representing temperatures of 40° C. andlower (scan 1705 of FIG. 17 corresponding to T=40° C.), at least onecharacteristic line 1770 is present that is indicative of the presenceof Form D. The scan 1710 representative of T=50° C. shows a diminishingof the line 1770. By T=60° C., the scan 1720 does not show the line 1770characteristic of Form D. At this point, the sample has an XRPD scanresembling Form A. Thus, Form D is also characterized by a transition toForm A below 60° C., and a loss of stability of Form D below 50° C. Theremaining scans show features at various temperatures that areindicative of Form A.

Form D is also characterized by the TGA measurement result shown in FIG.18; the measurement is performed with a heating rate of 5° C./min on asample containing Forms A and D. In particular, Form D is characterizedas being anhydrous since the first weight loss transition 1811 beginsabove the temperature at which the Form D crystals interconvert to FormA according to the temperature controlled XRPD measurements. Rather thefirst weight loss is corresponds with the loss of THF from the crystalsof the sample; this result is confirmed by simultaneous DSC and TGAcoupled to mass spectrometry analysis. A second weight loss transition1812 is observed with an interpolated beginning temperature of 164° C.,having a maximum weight loss rate at 180° C. This is similar to thetransition observed by TGA for Form A. Thus, the result confirms thetemperature-dependent XRPD result of the transformation of thecrystalline Form D fraction into Form A at high temperatures.

A DSC measurement is also used to characterize Form D as shown in FIG.19. The DSC measurement is performed on a mixture of Forms A and D ofthe Citrate Salt at a heating rate of 0.3° C./min. The curve 1910 showsan endothermic transition 1911 interpolated to begin at about 47° C. andhaving a T_(max)=52° C.; this transition is corresponds with theconversion of crystalline Form D to Form A, and thus characterizes thecrystalline form. The transition is also characterized by an integratedenthalpy of 3.7 J/g. The endothermic transition 1912, beginning at about60° C. and having a T_(max)=77° C., corresponds with the release of THFas discussed in the TGA analysis. The enthalpy of the transition isintegrated as 18.1 J/g. The endothermic transition 1913, interpolated tobegin at about 160° C. and having a T_(max)=166° C., corresponds withthe melting/decomposition of Form A.

Form D is characterized by a loss of stability, and interconversion toForm A, after exposure to changes in relative humidity. FIG. 20 displayscurves corresponding to four consecutive water sorption/desorptioncycles performed at 25° C. on a sample containing Forms A and D. Thecurves depict the percent change in mass, on a dry crystal basis, of thecrystal as a function of relative humidity. The sorption begins at 40%relative humidity, and is successively cycled between 100% relativehumidity and 0% relative humidity. A hysteresis effect is observed witheach humidification cycle, corresponding to a global weight loss ofabout 14.3% (w/w) after all 4 cycles.

XRPD measurements before and after the cycling show a structuralmodification in which the presence of Form D has disappeared after thecycling; only the signature of Form A remains.

Forms E and H

Form E is a single crystalline form of the Citrate Salt consistent withembodiments of the invention. A particular embodiment of the inventionis directed toward the single crystalline form having a substantiallysimilar XRPD pattern to what is displayed in FIG. 21. CuKα₁ radiation isused to generate the XRPD pattern. The single crystalline form isalternatively characterized by the first 30 lines of a XRPD pattern, aslisted in Table 5 (the d spacing values given in angstroms).

TABLE 5 Indexing of XRPD Pattern of FIG. 21 2θ λ_(Cu)K{circumflex over(α)} h k l d-spacing 1,54184 Å 1 0 0 13.522 6.54 0 0 1 12.464 7.09 −1 01 11.619 7.61 −1 1 0 8.256 10.72 0 1 1 7.997 11.06 1 0 1 7.808 11.33 −11 1 7.759 11.40 −2 0 1 7.195 12.30 −2 0 0 6.761 13.10 −1 0 2 6.709 13.201 1 1 6.249 14.17 0 0 2 6.232 14.21 −2 1 1 5.922 14.96 −2 0 2 5.80915.25 −2 1 0 5.673 15.62 −1 1 2 5.641 15.71 0 1 2 5.349 16.57 0 2 05.213 17.01 2 0 1 5.177 17.13 −2 1 2 5.075 17.48 1 0 2 4.987 17.79 −3 01 4.870 18.22 −1 2 0 4.864 18.24 0 2 1 4.809 18.45 −1 2 1 4.756 18.66 21 1 4.637 19.14 −3 0 2 4.565 19.44 3 0 0 4.507 19.70 1 1 2 4.499 19.74−1 0 3 4.476 19.83

Form E is also characterized by one or more of the peaks in Table 5.Embodiments of the invention utilize any number of the listed peaks inTable 5 including the option of using all of them. In another alternateembodiment of the invention, Form E is characterized by one or more ofthe calculated cell parameters of Table 6, derived from analysisconducted on a XRPD measurement. The crystalline structure is alsocharacterized to be orthorhombic P2₁ (Z=2 and Z′=1).

TABLE 6 Cell Parameters from XRPD Scan of Form E a = 14.612(3) Å b =10.425(1) Å c = 13.469(2) Å α = 90° β = 112.27(1)° γ = 90° V = 1898.8 Å³Z = 2 and Z′ = 1

Isothermal XRPD measurements, using CoKα₁ (λ=1.7890 Å) and CoKα₂(λ=1.7929 Å) radiation on a sample of Form E in a dry nitrogenenvironment, are presented in FIG. 22. The temperatures range fromambient temperature to 190° C. at intervals of 10° C.

In one embodiment of the invention, Form E is characterized by a loss ofstability between 50° C. and 60° C. Scan 2210, corresponding to the XRPDat 40° C., shows particular lines (e.g., line 2270) beginning todisappear, while other lines (e.g., line 2280) start to grow. At 60° C.(scan 2220), the structure characteristic of Form E has disappeared. Anew crystalline form of the Citrate Salt, designated Form H, is nowpresent. The scan 2230 at 120° C. shows the beginnings of thedestabilization of the crystalline Form H. The scan 2240 at 140° C.shows the that the sample has melted.

Thus, in another embodiment of the invention, Form H is characterized byeach of the scans 2290 of FIG. 22 corresponding to temperatures between60° C. and 130° C., and/or a loss of stability at a temperature between130° C. and 140° C. It may also be characterized by the transitionbetween Form E and Form H.

FIG. 23 shows the result of a TGA measurement on sample of crystallineForm E. The curve 2310 graphs the weight of the sample as a function oftemperature; corresponding curve 2320 graphs the derived rate of weightloss as a function of temperature.

Form E is characterized by the weight loss transition 2311, in terms ofthe interpolated beginning temperature 52° C., the maximum rate ofweight change temperature of 64° C., and/or the weight loss of 6.14%(w/w) associated with the transition. Form E is also characterized as ahydrated form of the Citrate Salt.

Form H of the Citrate Salt is characterized by the weight losstransition 2312, in terms of the interpolated beginning temperature of127° C., the maximum rate of weight change temperature of 142° C.,and/or the weight loss of 20.10% (w/w) associated with the transition.The transition 2311, as described above, also characterizes Form H.

FIG. 24 depicts the result of a DSC measurement on Form E of the CitrateSalt at a heating rate of 5° C./min. The curve 2410 graphs the heat flowinto the sample as a function of temperature.

Form E is characterized by the endothermic transition 2411 of the curve2410. The transition 2411 is characterized by an interpolated beginningtemperature of 45° C., with the maximum rate of heat loss at T_(max)=67°C.; the integrated enthalpy change is calculated to be 178.3 J/g. Thetransition 2411 corresponds with loss of water in the sample, and theconversion of Form E to Form H (i.e., the loss of stability in Form E).

Form H is characterized by another endothermic transition 2412 of thecurve 2410. The transition 2412 is characterized by an interpolatedbeginning temperature of 131° C., with the maximum rate of heat loss atT_(max)=141° C. The transition 2412 corresponds with the loss of waterfrom Form H, and the decomposition/melting of the crystal. Transition2411 also characterizes Form H.

FIG. 25 shows curves corresponding to two consecutive watersorption/desorption cycles performed at 25° C. on a sample of Form E.The curves, depicting the percent change in mass on a dry crystal basisof the crystal as a function of relative humidity, overlap each other.This indicates that the sorption phenomena is reversible. The curvesindicate the sample gains about 5.13% (w/w) as the relative humidity isincreased from 0% to 10%. Between 10% and 90% relative humidity, thesample weight changes about 2.28% (w/w). XRPD measurements show that thecrystal has a structure corresponding with Form E before thesorption/desorption cycles, and a structure corresponding with Form Hafter the cycling.

At ambient temperatures form E is stable from 10 to 90% relativehumidity. Under 10% relative humidity, and as experimentally observedwhen exposed at 0% relative humidity, form E fully transforms into formH. As indicated by sorption/desorption curves, form H reversiblytransforms into form E when exposed to 10% relative humidity or a higherrelative humidity rate.

Forms F and G

A crystalline containing a mixture of Forms F and G is characterized byany number of the lines identified by a * in the XRPD pattern of FIG.26, in an embodiment of the invention. FIG. 26 presents the relativeintensity as a function of 2θ for scans 3110, 3120 under dry nitrogenand ambient temperature using CoKα₁ (λ=1.7890 Å) and CoKα₂ (λ=1.7929 Å)radiation. Scan 2610 corresponds with a sample containing Forms E, F,and G. Scan 2620, corresponding with pure Form E, is used to distinguishthe peaks of scan 2610 that are unique to Forms F and G.

FIG. 27 records isothermal XRPD measurements using CoKα₁ (λ=1.7890 Å)and CoKα₂ (λ=1.7929 Å) radiation on a sample containing Forms E, F, andG at temperatures from ambient to 210° C. in steps of 10° C. Each curverepresents an isothermal scan of the sample in a dry nitrogenenvironment.

Form E is characterized by a loss of stability at temperatures above 50°C. The scan 2710, representing the measurement at 50° C., indicates thatlines 2711 unique to Form E have disappeared at temperatures of 50° C.and higher. Scans with temperatures of 110° C. and higher show thatlines unique to Form F (e.g., line 2721) have disappeared. Thus, Form Fis characterized by a loss of stability of this form at temperaturesgreater than 110° C. (scan 2720 showing the measurement at 110° C.).Form G is characterized by a loss of stability at temperatures of 130°C. and higher; scans at temperatures of 130° C. 2730 and higher showlines corresponding with Form G have disappeared (e.g., line 2731).

Throughout the measurements as temperatures increase from ambient to130° C., lines corresponding with Form B continue to grow in the variousscans. At temperatures 130° C. and higher, the scans correspondsubstantially with the lines for Form B. Thus, the data from these scansindicates that each of Forms E, F, and G converts to Form B as thetemperature is raised.

A TGA measurement on a sample of containing Form E, F, and G is shown inFIG. 28. The curve 2810 records the weight of the sample as a functionof temperature. Curve 2820 records the corresponding derived rate ofweight loss as a function of temperature for a sample heated at 5°C./min. Four weight losses are observed in FIG. 28; the relevant datasummarized in Table 7.

TABLE 7 Weight Transition Data for a TGA Measurement of a SampleContaining Form E, F, and G T (° C.) corresponding to Weight WeightTransition Interpolated Max. Rate of Weight Loss % (label in FIG. 28)Starting T (° C.) Change (w/w) 2811 42 50 1.9 2812 110 123 3.6 2813 131138 1.6 2814 166 187 22.6

Form E is characterized by the weight loss 2811, which corresponds withthe conversion away from Form E. Form F is characterized by the weightloss 2812, which corresponds with the conversation away from Form F.Form G is characterized by the weight loss 2813, which corresponds withthe conversion away from Form G. The fourth weight loss 2814 is thedecomposition/melting of crystalline Form B, and the corresponding lossof citrate.

FIG. 29 presents the result of a DSC measurement on sample containingForms E, F, and G at a heating rate of 5° C./min. The curve 2910 showsfour endothermic transitions, the quantitative data summarized in Table8.

TABLE 8 Heat Flow Transition Data for a DSC Measurement of a SampleContaining Form E, F, and G Transition (label Interpolated IntegratedEnthalpy in FIG. 34) Starting T (° C.) T_(max) (° C.) Change (J/g) 341178 99 20.3 3412 119 124 5.8 3413 131 136 2.4 3414 165 181 N/A

The first endothermic transition 2911 characterizes the conversion ofcrystals with Form E to Form B. The second endothermic transition 2912characterizes the conversion of crystals with Form F to Form B. Thethird endothermic transition 2913 characterizes the conversion ofcrystals with Form G to Form B. The fourth endothermic transition 2914characterizes the decomposition/melting of crystalline Form B.

FIG. 30 displays curves corresponding to four consecutive watersorption/desorption cycles performed at 25° C. on a sample containingForms E, F, and G. The graphs depict the percent change in mass, on adry crystal basis, of the crystal as a function of relative humidity.The first sorption run begins at 40% relative humidity, and issuccessively cycled between 100% relative humidity and 0% relativehumidity. A hysteresis effect is observed for the firstsorption/desorption cycle; the remaining 3 cycles being reversible andnot displaying hysteresis. The net weight loss in the sample aftersorption/desorption cycling with water is about 7.5% (w/w), which issimilar to the total of the three weight transitions observed from theTGA analysis of FIG. 28 at the lower temperatures. Thus, heating thecycled sample to a temperature of about 131° C. or higher will convertthe sample to anhydrous Form B.

Subjecting the sample to a change in relative humidity from 0% to 10%results in a water uptake corresponding to 3.76% (w/w). A change inrelative humidity from 10% to 90% results in a water uptakecorresponding to 2.10% (w/w). A change in relative humidity from 90% to100% results in a water uptake corresponding between about 0.82% (w/w)to about 0.93% (w/w). These changes are all reversible.

XRPD measurements taken of the sample at 40% relative humidity beforeand after the sorption/desorption cycling show that Form F disappearsand the lines of Form G have dissipated, while Form E lines grow, afterthe cycling is performed. The sorption/desorption weight changes between10% and 90% relative humidity are similar to what is witnessed for FormE (c.f. FIG. 25). Thus, humidification cycling results in the conversionof Forms F and G to Form E.

The same change of the loss of lines of Form F and diminution of linesof Form G, while the lines of Form E grow, is witnessed when comparingXRPD of batch F1 before and after the sample is subjected to a relativehumidity of 97.5% for about one month, or before and after the sample isrinsed with water.

XRPD measurements made on a sample of batch F1 subject to a relativehumidity of 0% for about one month show a mixture of Forms E and B;total conversion to Form B is not observed.

Other embodiments of the invention are directed to a single crystallineform of the Citrate Salt characterized by a combination of theaforementioned characteristics of any of the single crystalline formsdiscussed herein. The characterization may be by any combination of oneor more of the XRPD, TGA, DSC, and water sorption/desorptionmeasurements described for a particular polymorph. For example, thesingle crystalline form of the Citrate Salt may be characterized by anycombination of the XRPD results regarding the 2θ position of the majorpeaks in a XRPD scan; any combination of one or more of the cellparameters derived from a XRPD scan; and the temperature at which acrystalline form begins to destabilize, or decomposes/melts, asdetermined from XRPD scans of a sample of the crystalline form taken atdifferent temperatures. The single crystalline form of the Citrate Saltmay also be characterized by TGA determinations of the temperature atwhich a sample begins to undergo a weight loss transition and/or thetemperature corresponding with the rate of maximum weight change duringa weight loss transition. DSC determinations of T_(max) and/or thetemperature at which a sample begins to undergo a heat flow transitionmay also characterize the crystalline form. Weight change in a sampleand/or change in sorption/desorption of water per molecule of anhydrousCitrate Salt as determined by water sorption/desorption measurementsover a range of relative humidity (e.g., 0% to 90%) may alsocharacterize a single crystalline form of the Citrate Salt.

Examples of combinations of single crystalline form characterizationsusing multiple analytical techniques include the 2θ location of at leastone of the major peaks of a XRPD scan and the T_(max) in a heat flowtransition/endothermic transition observed by a corresponding DSCmeasurement; the 2θ location of at least one of the major peaks of aXRPD scan and the temperature at which a maximum rate of weight changeis noticed for a weight loss transition in a corresponding TGAmeasurement; the 2θ location of at least one of the major peaks of aXRPD scan, the T_(max) in a heat flow transition/endothermic transitionobserved by a corresponding DSC measurement, and the temperature atwhich a maximum rate of weight change is noticed for a weight losstransition in a corresponding TGA measurement; and the 2θ location of atleast one of the major peaks of a XRPD scan, the T_(max) in a heat flowtransition/endothermic transition observed by a corresponding DSCmeasurement, the temperature at which a maximum rate of weight change isnoticed for a weight loss transition in a corresponding TGA measurement,and the change in sorption/desorption of water per molecule of anhydroussalt as determined by water sorption/desorption measurements over arange of relative humidity. As well, each of the aforementioned examplesmay replace the use of the 2θ location of at least one of major peaks ofa XRPD scan with one or more cell parameters of the single crystallineform, as determined from a XRPD scan, in consistent embodiments of theinvention.

The combinations of characterizations that are discussed above may beused to describe any of the polymorphs of the Citrate Salt discussedherein (e.g., Form A, B, C, D, E, F, G, or H). In an alternativeembodiment of the invention, a crystalline Citrate Salt, or a singlecrystalline form of the Citrate Salt, is characterized by the lack ofone or more properties associated with a particular polymorph (e.g., notbeing Form A, not having a T_(max)=167° C. when measured by DSC with aheating rate of 1° C./min, not having at least one of the major peaks ofForm A as measured by XRPD). In another alternative embodiment of theinvention, a crystalline Citrate Salt is characterized using acombination of the single crystalline form characterizations ofdifferent polymorphs (e.g., a crystalline form comprising Form A andForm B; a crystalline form characterized by have at least one of themajor peaks listed for Form A and Form B from XRPD measurements; thecrystalline form is anhydrous). Other alternative embodiments of theinvention relate to a crystalline Citrate Salt characterized by nothaving a combination of single crystalline form characterizations ofdifferent polymorphs (e.g., a crystalline Citrate Salt not having themajor peaks of XRPD patterns associated with Forms A, C, and D).

Certain other exemplary embodiments are described directly below.

In some embodiments, at least 10% by weight of the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol(“Citrate Salt”) is crystalline. In other embodiments, at least 70% byweight of the Citrate Salt is crystalline. In still other embodiments,at least 90% by weight of the Citrate Salt is crystalline.

In other embodiments, at least 10% by weight of the Citrate Salt is asingle crystalline form. In yet other embodiments, at least 70% byweight of the Citrate Salt is a single crystalline form. In still otherembodiments, at least 90% by weight of the Citrate Salt is a singlecrystalline form.

In other embodiments, at least 10% by weight of the Citrate Salt isother than Form A. In still other embodiments, at least 70% by weight ofthe Citrate Salt is other than Form A. In yet other embodiments, atleast 90% by weight of the Citrate Salt is other than Form A.

In other embodiments, a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olis provided, the citrate salt being substantially Form A. In still otherembodiments, a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olis provided the citrate salt being at least 10% by weight Form A. In yetother embodiments, a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olis provided, the citrate salt being at least 70% by weight Form A.

In still other embodiments, the Citrate Salt is at least 70% by weight asingle crystalline form and the crystalline form is characterized by atleast one of the x-ray powder diffraction peaks at 2θ angles of 9.8,11.7, 12.6, 15.5, 15.7, 15.9, 17.3, 17.5, 18.2, 19.0, and 19.7, whenmeasured with CuKα₁ radiation. In other embodiments, the Citrate Salt isat least 70% by weight a single crystalline form and the crystallineform is characterized by x-ray powder diffraction peaks at 2θ angles of9.8, 11.7, 12.6, 15.5, 15.7, 15.9, 17.3, 17.5, 18.2, 19.0, and 19.7,when measured with CuKα₁ radiation. In still other embodiments, thesingle crystalline form is characterized by a x-ray powder diffractionpattern substantially similar to FIG. 1 when measured with CuKα₁radiation. In yet other embodiments, the single crystalline form ischaracterized by having a stability transition in the range of 150° C.and 160° C. as observed with controlled temperature x-ray powderdiffraction.

In other embodiments, at least 70% by weight of the Citrate Salt is asingle crystalline form, wherein the single crystalline form ischaracterized by a T_(max) of 167° C.±3° C. during an endothermictransition observed by differential scanning calorimetry using ascanning rate of 1° C./minute. In still other embodiments, at least 70%by weight of the Citrate Salt is a single crystalline form, wherein thesingle crystalline form is characterized by a weight loss observed bythermal gravimetric analysis corresponding to decomposition of thecitrate salt beginning at 164° C.±3° C. In still other embodiments, atleast 70% by weight of the Citrate Salt is a single crystalline form,wherein the single crystalline form gains about 0.8 moles of water permole of citrate salt over a relative humidity change from 0% to 90% at25° C.

In still other embodiments, a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olis provided, the citrate salt being substantially Form B. In yet otherembodiments, a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olis provided, the citrate salt being at least 10% by weight Form B. Instill other embodiments, at least 70% by weight of the Citrate Salt is asingle crystalline form, wherein the single crystalline form is Form B.In yet other embodiments, at least 70% by weight of the Citrate Salt isa single crystalline form, wherein the single crystalline form ischaracterized by at least one of the x-ray powder diffraction peaks at2θ angles of 10.6, 11.6, 12.3, 14.8, 15.8, 16.1, 16.7, 18.8, 20.6, 21.7,and 24.5, when measured with CuKα₁ radiation. In yet other embodiments,at least 70% by weight of the Citrate Salt is a single crystalline form,wherein the single crystalline form is characterized by x-ray powderdiffraction peaks at 2θ angles of 10.6, 11.6, 12.3, 14.8, 15.8, 16.1,16.7, 18.8, 20.6, 21.7, and 24.5, when measured with CuKα₁ radiation. Instill other embodiments, the single crystalline form is characterized bya x-ray powder diffraction pattern substantially similar to FIG. 6 whenmeasured with CuKα₁ radiation. In yet other embodiments, the singlecrystalline form is characterized by having a stability transition inthe range of 170° C. and 180° C. as observed with controlled temperaturex-ray powder diffraction.

In yet other embodiments, at least 70% by weight of the Citrate Salt isa single crystalline form, wherein the single crystalline form ischaracterized by a T_(max) of 179° C.±3° C. during an endothermictransition observed by differential scanning calorimetry using ascanning rate of 1° C./minute. In yet other embodiments, at least 70% byweight of the Citrate Salt is a single crystalline form, wherein thesingle crystalline form is characterized by a weight loss observed bythermal gravimetric analysis corresponding to decomposition of thecitrate salt beginning at 180° C.±3° C. In yet other embodiments, atleast 70% by weight of the Citrate Salt is a single crystalline form,wherein the single crystalline form gains about 0.4 moles of water permole of citrate salt over a relative humidity change from 0% to 90% at25° C.

Pharmaceutical Compositions

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or diluent;and a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olas discussed herein. In some embodiments, the Citrate Salt issubstantially crystalline.

The present invention also provides a pharmaceutical compositioncomprising pharmaceutically acceptable carrier or diluent; and citratesalt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,wherein at least 10% by weight of the citrate salt is crystalline. Insome embodiments, at least 10% by weight of the citrate salt is a singlecrystalline form. In still other embodiments, at least 10% by weight ofthe citrate salt is other than Form A.

The present invention further provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or diluent; and acitrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,wherein at least 70% by weight of the citrate salt is crystalline. Insome embodiments, at least 70% by weight of the citrate salt is a singlecrystalline form. In other embodiments, at least 70% by weight of thecitrate salt is other than Form A. In yet other embodiments, the singlecrystalline form is Form A. In still other embodiments, the singlecrystalline form is characterized by at least one of the x-ray powderdiffraction peaks at 2θ angles of 9.8, 11.7, 12.6, 15.5, 15.7, 15.9,17.3, 17.5, 18.2, 19.0, and 19.7, when measured with CuKα₁ radiation. Inyet other embodiments, the single crystalline form is characterized byx-ray powder diffraction peaks at 2θ angles of 9.8, 11.7, 12.6, 15.5,15.7, 15.9, 17.3, 17.5, 18.2, 19.0, and 19.7, when measured with CuKα₁radiation. In still other embodiments, the single crystalline form ischaracterized by a x-ray powder diffraction pattern substantiallysimilar to FIG. 1 when measured with CuKα₁ radiation. In yet otherembodiments, the single crystalline form is characterized by having astability transition in the range of 150° C. and 160° C. as observedwith controlled temperature x-ray powder diffraction. In still otherembodiments, the single crystalline form is characterized by a T_(max)of 167±3° C. during an endothermic transition observed by differentialscanning calorimetry using a scanning rate of 1° C./minute. In yet otherembodiments, the single crystalline form is characterized by a weightloss observed by thermal gravimetric analysis corresponding todecomposition of the citrate salt beginning at 164° C.±3° C. In stillother embodiments, single crystalline form gains about 0.8 moles ofwater per mole of citrate salt over a relative humidity change from 0%to 90% at 25° C. In still other embodiments, the single crystalline formis Form B. In yet other embodiments, the single crystalline form ischaracterized by at least one of the x-ray powder diffraction peaks at20 angles of 10.6, 11.6, 12.3, 14.8, 15.8, 16.1, 16.7, 18.8, 20.6, 21.7,and 24.5, when measured with CuKα₁ radiation. In still otherembodiments, the single crystalline form is characterized by x-raypowder diffraction peaks at 2θ angles of 10.6, 11.6, 12.3, 14.8, 15.8,16.1, 16.7, 18.8, 20.6, 21.7, and 24.5, when measured with CuKα₁radiation. In still other embodiments, the single crystalline form ischaracterized by a x-ray powder diffraction pattern substantiallysimilar to FIG. 6 when measured with CuKα₁ radiation. In yet otherembodiments, the single crystalline form is characterized by having astability transition between 170° C. and 180° C. as observed withcontrolled temperature x-ray powder diffraction. In still otherembodiments, the single crystalline form is characterized by a T_(max)of 179° C.±3° C. during an endothermic transition observed bydifferential scanning calorimetry using a scanning rate of 1° C./minute.In yet other embodiments, the single crystalline form is characterizedby a weight loss observed by thermal gravimetric analysis correspondingto decomposition of the citrate salt beginning at 180° C.±3° C. In stillother embodiments, the single crystalline form gains about 0.4 moles ofwater per mole of citrate salt over a relative humidity change from 0%to 90% at 25° C.

The present invention also provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or diluent; and acitrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,wherein at least 90% by weight of the citrate salt is crystalline. Insome embodiments, at least 90% by weight of the citrate salt is a singlecrystalline form. In still other embodiments, at least 90% by weight ofthe citrate salt is other than Form A.

The present invention further provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or diluent; and acitrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,the citrate salt being substantially Form A.

In yet other embodiments, the present invention also provides apharmaceutical composition comprising a pharmaceutically acceptablecarrier or diluent; and a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,the citrate salt being at least 10% by weight Form A.

In still other embodiments, the present invention provides apharmaceutical composition comprising a pharmaceutically acceptablecarrier or diluent; and a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,the citrate salt being substantially Form B. In yet other embodiments,the present invention provides a pharmaceutical composition comprising apharmaceutically acceptable carrier or diluent; and a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,the citrate salt being at least 10% by weight Form B.

As described above, the pharmaceutical compositions of the presentinvention additionally comprise a pharmaceutically acceptable carrier,which, as used herein, includes any and all solvents, diluents, or otherliquid vehicle, dispersion or suspension aids, surface active agents,isotonic agents, thickening or emulsifying agents, preservatives, solidbinders, lubricants and the like, as suited to the particular dosageform desired. Remington's Pharmaceutical Sciences, Mack Publishing Co.,a standard reference text in this field, discloses various carriers usedin formulating pharmaceutical compositions and known techniques for thepreparation thereof. Except insofar as any conventional carrier mediumis incompatible with the Citrate Salt, such as by producing anyundesirable biological effect or otherwise interacting in a deleteriousmanner with any other component(s) of the pharmaceutical composition,its use is contemplated to be within the scope of this invention.

The pharmaceutical compositions of the invention can be manufactured bymethods well known in the art such as conventional granulating, mixing,dissolving, encapsulating, lyophilizing, or emulsifying processes, amongothers. Compositions may be produced in various forms, includinggranules, precipitates, or particulates, powders, including freezedried, rotary dried or spray dried powders, amorphous powders, tablets,capsules, syrup, suppositories, injections, emulsions, elixirs,suspensions or solutions. Formulations may optionally containstabilizers, pH modifiers, surfactants, bioavailability modifiers andcombinations of these.

The quantity of active ingredient in the composition can range fromabout 0.1% to about 99.9% by weight, or about 20% to about 80% byweight. A unit dose preparation can contain from 1 mg to about 1000 mgactive ingredient, preferably about 10 mg to about 100 mg activeingredient. The composition can, if desired, also contain othercompatible therapeutic agents, including but not limited to,theophylline, β-adrenergic bronchodilators, corticosteroids,antihistamines, antiallergic agents, immunosuppressive agents (e.g.,cyclosporine A, FK-506, prednisone, methylprednisolone), hormones (e.g.,adrenocorticotropic hormone (ACTH)), cytokines (e.g., interferons (e.g.,INFβ-1a, INFβ-1b)) and the like.

Treatment with Citrate Salt and Crystalline Forms Thereof

Another aspect of the invention relates to a method of treatment,including prophylactic and therapeutic treatments, of a diseaseassociated with aberrant leukocyte recruitment and/or activation ormediated by chemokines or chemokine receptor function, including chronicinflammatory disorders characterized by the presence of RANTES, MIP-1α,MCP-2, MCP-3 and/or MCP-4 responsive T cells, monocytes and/oreosinophils, including but not limited to diseases such as arthritis(e.g., rheumatoid arthritis), atherosclerosis, arteriosclerosis,restenosis, ischemia/reperfusion injury, diabetes mellitus (e.g., type 1diabetes mellitus), psoriasis, multiple sclerosis, inflammatory boweldiseases such as ulcerative colitis and Crohn's disease, rejection oftransplanted organs and tissues (i.e., acute allograft rejection,chronic allograft rejection), graft versus host disease, chronicobstructive pulmonary disorder (COPD), as well as allergies and asthma.Other diseases associated with aberrant leukocyte recruitment and/oractivation which can be treated (including prophylactic treatments) withthe methods disclosed herein are inflammatory diseases associated withHuman Immunodeficiency Virus (HIV) infection. Still other diseasesassociated with aberrant leukocyte recruitment and/or activation whichcan be treated (including prophylactic treatments) with the methodsdisclosed herein are cancer and osteolytic bone disorders. The methodcomprises administering to the subject in need of treatment an effectiveamount of a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-azadibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,as discussed herein. In some embodiments, the Citrate Salt issubstantially crystalline. In other embodiments, at least 10% by weightof the Citrate Salt is crystalline. In yet other embodiments, at least70% by weight of the citrate salt is at crystalline. In still otherembodiments, at least 70% by weight of the Citrate Salt is a singlecrystalline form. In other embodiments, the single crystalline form isForm A. In still other embodiments, the single crystalline form is FormB. In still other embodiments, at least 70% by weight of the citratesalt is other than Form A.

In other embodiments, the method is useful for the treatment ofrheumatoid arthritis, multiple sclerosis, atherosclerosis, inflammatorybowel disease, or psoriasis. In still other embodiments, the method isuseful for the treatment of arteriosclerosis, restenosis, diabetesmellitus, colitis, or Crohn's disease. In yet other embodiments, themethod is useful for the treatment of rejection of transplanted organs,graft versus host disease, allergies, asthma, or an inflammatory diseaseassociated with Human Immunodeficiency Virus infection. In still otherembodiments, the method is useful for the treatment of chronicobstructive pulmonary disorder (COPD).

In other embodiments the present invention provides a method fortreating rheumatoid arthritis comprising administering to a subject inneed thereof an effective amount of a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.In some embodiments, the Citrate Salt used for the treatment ofrheumatoid arthritis has any one or more of the properties describedabove and herein for the Citrate Salt. In other embodiments, the CitrateSalt used for the treatment of rheumatoid arthritis has one or more ofthe following properties:

-   -   1. the Citrate Salt is substantially crystalline;    -   2. at least 70% by weight of the Citrate Salt is a single        crystalline form;    -   3. at least 70% by weight of the Citrate Salt is a single        crystalline form and the single crystalline form is Form A or        Form B;    -   4. at least 70% by weight of the Citrate Salt is other than Form        A.

In other embodiments, the present invention provides a method fortreating multiple sclerosis comprising administering to a subject inneed thereof an effective amount of a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.In some embodiments, the Citrate Salt used for the treatment of multiplesclerosis has any one or more of the properties described above andherein for the Citrate Salt. In other embodiments, the Citrate Salt usedfor the treatment of multiple sclerosis has one or more of the followingproperties:

-   -   1. the Citrate Salt is substantially crystalline;    -   2. at least 70% by weight of the Citrate Salt is a single        crystalline form;    -   3. at least 70% by weight of the Citrate Salt is a single        crystalline form and the single crystalline form is Form A or        Form B;    -   4. at least 70% by weight of the Citrate Salt is other than Form        A.

In yet other embodiments, the present invention provides a method fortreating chronic obstructive pulmonary disorder (COPD) comprisingadministering to a subject in need thereof an effective amount of acitrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.In some embodiments, the Citrate Salt used for the treatment of chronicobstructive pulmonary disorder has any one or more of the propertiesdescribed above and herein for the Citrate Salt. In other embodiments,the Citrate Salt used for the treatment of multiple sclerosis has one ormore of the following properties:

-   -   1. the Citrate Salt is substantially crystalline;    -   2. at least 70% by weight of the Citrate Salt is a single        crystalline form;    -   3. at least 70% by weight of the Citrate Salt is a single        crystalline form and the single crystalline form is Form A or        Form B;    -   4. at least 70% by weight of the Citrate Salt is other than Form        A.

Other embodiments of the invention relate to methods of antagonizing achemokine receptor, such as CCR1, in a subject comprising administeringto the mammal the Citrate Salt as described herein.

According to the method, chemokine-mediated chemotaxis and/or activationof pro-inflammatory cells bearing receptors for chemokines can beinhibited. The amount of the Citrate Salt administered to the individualwill depend on the type and severity of the disease and on thecharacteristics of the individual, such as general health, age, sex,body weight and tolerance to drugs. It will also depend on the degree,severity and type of disease. The skilled artisan will be able todetermine appropriate dosages depending on these and other factors.Typically, an effective amount of the compound can range from about 0.1mg per day to about 100 mg per day for an adult. Preferably, the dosageranges from about 1 mg per day to about 100 mg per day. An antagonist ofchemokine receptor function can also be administered in combination withone or more additional therapeutic agents, e.g. theophylline,β-adrenergic bronchodilators, corticosteroids, antihistamines,antiallergic agents, immunosuppressive agents (e.g., cyclosporin A,FK-506, prednisone, methylprednisolone), hormones (e.g.,adrenocorticotropic hormone (ACTH)), cytokines (e.g., interferons (e.g.,IFNβ-1a, IFNβ-1b)) and the like.

The Citrate Salt can be administered by any suitable route, including,for example, orally in capsules, suspensions or tablets or by parenteraladministration. Parenteral administration can include, for example,systemic administration, such as by intramuscular, intravenous,subcutaneous, or intraperitoneal injection. The Citrate Salt can also beadministered orally (e.g., dietary), transdermally, topically, byinhalation (e.g., intrabronchial, intranasal, oral inhalation orintranasal drops), or rectally, depending on the disease or condition tobe treated. Oral or parenteral administration are preferred modes ofadministration.

The Citrate Salt can be administered to the individual in conjunctionwith an acceptable pharmaceutical or physiological carrier as part of apharmaceutical composition for treatment the diseases discussed above.Formulation of a compound to be administered will vary according to theroute of administration selected (e.g., solution, emulsion, capsule).

Preparation of Crystal Forms of the Citrate Salt

Some embodiments of the invention are directed toward a process ofpreparing a crystalline citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.In one embodiment, citric acid is combined with a solution of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olin a solvent to precipitate the crystalline citrate salt. Thecrystalline citrate salt is then isolated from the mixture usingtechniques known to those skilled in the art (e.g., filtration,evaporation, decanting, distillation (vacuum or at atmosphericpressure)).

Solvents that may be used to crystallize the Citrate Salt includeethanol, acetone, methanol, heptane, methyl ethyl ketone (MEK), water,toluene, isopropanol, n-propanol, tetrahydrofuran (THF), acetonitrile,dimethylsulfoxide (DMSO), dichlormethane, and combinations of one ormore of the aforementioned solvents. The resulting crystalline CitrateSalt may include one or more single crystalline forms.

In particular embodiments, solvents such as acetone or toluene may beused to crystallize Form A of the Citrate Salt.

In other particular embodiments, solvents such as ethanol,methanol/heptane, MEK/water, n-propanol, isopropanol, isopropanol/water,acetonitrile/water, or methanol may be used to crystallize Form B. Moreparticularly, Form B may be created by taking samples of Form A andexposing them to one of the solvent systems listed above. The particularconditions of the exposure are listed in Table 9.

In another particular embodiment of the invention, a mixture of Forms Aand C is prepared by dissolving a sample of Form A in a mixture of THFand water at high temperature, and subsequently crystallizing themixture of forms. Alternatively, a mixture of Forms A and C is preparedby exposing a sample of Forms A and B to THF at ambient temperature, andletting the crystal mature to Forms A and C.

In another particular embodiment of the invention, a mixture of Forms Aand D is prepared by dissolving a sample of Form A in THF and slowlyevaporating the solvent at ambient temperature and atmospheric pressure.

A particular embodiment of the invention is directed to preparing Form Eby exposing a sample of Form A to water at ambient temperature, andallowing the sample to mature to Form E.

An alternate embodiment of the invention is directed to forming acrystalline Citrate Salt including Forms E, F, and G. A sample of theamorphous form of the Citrate Salt is exposed to water at ambienttemperature. The sample subsequently matures to include a mixture ofForms E, F, and G.

A preferred embodiment of the invention is a process of preparing Form Aof the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.The process comprises combining citric acid and a solution of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olin acetone to precipitate the crystalline citrate salt. The salt issubsequently isolated from the combination. The mixture is typicallyheld at a temperature between 30° C. and 40° C. during theprecipitation, but other suitable temperatures can also be used. Thetemperature may be held for at least 10 minutes. Typically, at least oneequivalent of citric acid is used per equivalent of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol;preferably 1.0 to 1.5 equivalents of the citric acid are used.

In another preferred embodiment of the invention, a process forpreparing Form B of the Citrate Salt includes combining citric acid withthe(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olin ethanol to precipitate the crystalline citrate salt, and isolatingthe salt subsequently. Processes in accord with embodiments of theinvention may further include seeding the mixture for precipitation withcrystals of the Citrate Salt. In a preferred related embodiment, thecrystals are Form B of the Citrate Salt. The process is typicallyconducted between 18° C. and 22° C. while combining the elements of themixture; other suitable temperatures may also be utilized. Thetemperature is typically held for at least 2 hours while the crystalsprecipitate. Typically, at least one equivalent of citric acid is usedper equivalent of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.

Some embodiments of the invention are directed toward a mixture forcrystallizing a citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.The mixture comprises citric acid;(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol;and a crystallization medium.

A crystallization medium is a solvent or a solvent system (e.g.,combination of solvents) in which the citric acid and(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxo-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olare soluble, but the crystalline Citrate Salt is insoluble or at mostsparingly soluble. The solubility of the(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-oland Citrate Salt depends upon the temperature of the mixture (i.e., thesolubility being higher at higher temperature).

Examples of crystallization mediums include ethanol, acetone, methanol,heptane, methyl ethyl ketone (MEK), water, toluene, isopropanol,n-propanol, tetrahydrofuran (THF), acetonitrile, dimethylsulfoxide(DMSO), dichlormethane, and combinations of one or more of theaforementioned solvents. In a particular embodiment of the invention,the crystallization medium is ethanol; acetone; methanol; MEK;n-propanol; isopropanol; THF; toluene; acetonitrile; water; heptane; amixture of methanol and heptane; a mixture of MEK and water; a mixtureof acetonitrile and water; a mixture of isopropanol and water; or amixture of THF and water. In another particular embodiment of theinvention, the crystallization medium is acetone. In yet anotherparticular embodiment of the invention, the crystallization medium isethanol. In still another particular embodiment of the invention, thecrystallization medium is any of the solvents/solvent systems used inTable 9 herein.

Mixtures for crystallizing a Citrate Salt may include one or more seedsof a crystal that are optionally slurried in a medium; aprecipitation/crystallization aid; and/or being held at various times atvarious temperatures to aid crystallization of the Citrate Salt. Otheradditives, such as those used to enhance the rate of crystallization asrecognized by those of ordinary skill, are also within the scope of theinvention.

Though embodiments of the invention described herein refer specificallyto the (S)-enantiomer of the compound4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,it is well understood to those of ordinary skill in the art that relatedembodiments of the invention include use of the (R)-enantiomer, aracemic mixture of the compound, and any fractional combination of the(R) and (S) enantiomers.

Experimental Analytical Characterization of Samples of Crystalline Formsof the Citrate Salt

Crystalline forms of the Citrate Salt are characterized with the resultsfrom operating one or more analytical techniques on a sample of thecrystalline form. Such techniques include various types of X-ray powderdiffraction (XRPD), thermal gravimetric analysis (TGA), differentialscanning calorimetry (DSC), dynamic vapor sorption measurements (DVS),and optical microscopy. For each of the polymorphs of the Citrate Salt,in accord with an embodiment of the invention, one or more of theseanalytical techniques are utilized to characterize the polymorph. Theindividual techniques are described below.

X-ray powder diffraction (XRPD) is performed on various samples of theCitrate Salt using one of three instruments. XRPD measurements withInstrument 1 utilize a Siemens-Bruker D5000 Matic powder diffractometerwith a Bragg-Brentano (vertical θ-2θ configuration) parafocusinggeometry. X-rays are produced from a sealed cobalt anode running at 40kV and 30 mA. Two lines are typically emitted: CoKα₁ (λ=1.7890 Å) andCoKα₂ (λ=1.7929 Å). An iron β-filter limits the CoKβ (λ=1.6208 Å)radiation to about 1% of the diffracted beam at the detector. Theprimary beam passes through a parallel plate collimator (0.2 mm Sollerslits), then through a divergence slit (0.2 mm). Diffracted X-rays aredetected with a Braun 50 M multichannel linear detector having a10°-wide detection window in angle 2θ. Scans are performed from 1.5° to50.0° in 2θ, the scan time being 10 to 40 seconds per degree in 2θ. Theerror in the 2θ locations are typically within ±0.10 degrees.

In Instrument 2, high resolution XRPD diagrams are obtained with aPhilips Analytical X'Pert Pro MPD powder diffractometer utilizing theBragg-Brentano (vertical θ-2θ configuration) parafocusing geometrycoupled with a X'Celerator detector. A sealed copper anode X-ray tube,running at 45 kV and 40 mA, generates the impinging X-rays. An incidentbeam monochromator (Johansson type: a symmetrically cut curved germanium(111) crystal) produces pure CuKα₁ radiation (λ=1.54060 Å). A thin layerof the product is deposited on a single-crystal silicon wafer, cut outaccording to Si(510) crystallographic orientation that, by systematicextinction, impedes any Bragg reflection. In order to bring morecrystallites into the diffraction position, and thus reduce theinfluence of particle statistics on the measurements, a sample spinneris used. The spinner rotation speed is set at 1 revolution per second.The angular range extends from 3° to 40° or 50° in 2θ, with a 0.02° stepsize in 2θ. A counting time of 1250 to 3500 seconds per step is used.The error in the 2θ locations are typically within ±0.10 degrees.

Scans performed with Instrument 2 may be analyzed with the “ProfileFitting” module of the Diffrac-AT program furnished by the BrukerCorporation to determine the 2θ angular position of each of theobservable lines. Comparison of full widths at half maximum of the linesprovided by the module resolves peak overlapping, and reveals thepresence or absence of another phase. The best solution, the highestfigure of merit, given by the Accelrys company's “X-Cell” indexingprogram is subsequently optimized by Pawley refinement. The treatmentaccounts for the whole profile of the diagram, and not simply theposition of the lines. The treatment seeks to reproduce the experimentaldiagram as closely as possible by utilizing the diffracted intensitiesas variables.

In Instrument 3, XRPD measurements are performed at differenttemperatures to elucidate the evolution of crystal structure withtemperature. Tests are carried out with a Siemens-Bruker D5000diffractometer equipped with the Bragg Brentano parafocusing (θ-θ)geometry and an Anton-Paar TTK temperature chamber. Measurements utilizedry nitrogen or nitrogen streams having a particular humidity. Theinstrument specifications are substantially similar to theSiemens-Bruker D5000 Matic instrument described above (Instrument 1).Temperature is allowed to rise at a rate of 0.05° C./sec. Scans arerecorded under the following conditions: a 1.5 to 50.0 degree scan inangle 2θ, 10 to 20 seconds counting time per degree in 2θ. Data areacquired in isotherm mode when the requested temperature is reached.

Measurements utilizing thermal gravimetric analysis (TGA) are carriedout on a T.A. instruments TGAQ500 analyzer. Mass calibration isperformed with 10 and 100 mg certified masses. Temperature calibrationis performed with Alumel® and nickel standards (Curie points ofrespectively 154° C. and 354° C.). Samples are exposed to a constantnitrogen stream of 60 ml/min, and temperature ranges from roomtemperature to 350° C. at a 5° C./min rate. The quantity of producttested is between 2 and 13 mg. The sample is deposited in an openaluminum sample pan, which is itself placed in a platinum pan. Thetemperatures identified from TGA scans of a particular sample areaccurate to within ±3° C.

Differential scanning calorimetry (DSC) analyses are performed using aT.A Instruments Q1000 thermal analyser. Analyses are carried out usingnitrogen purge gas flowing at an average rate of 50 ml/min. A mechanicalcompressor cools the system, and equilibrates the instruments being usedat ambient temperature between analyses. The calorimeter is temperaturecalibrated with materials such as indium and lead (onset of meltingtemperatures of 156.6° C. and 327.5° C. respectively). Energycalibration is performed with a certified indium calibrator (meltingenthalpy of 28.45 J/g), applying a heating rate of 5° C./min. Thetemperatures identified from DSC scans of a particular sample areaccurate to within ±3° C.

The samples are submitted to the following experimental temperatureprograms: after the equilibration of the sample at 10° C., the specimenswere heated from 10° C. to 210° C. (or 250° C. or 260° C.) at a rate of1 to 50° C./min.

Modulated temperature differential scanning calorimetry (MTDSC) is alsoused since deconvolution of the total heat flow into reversing andnon-reversing heat flows allows the transitions to be easily identified.The samples were submitted to the same experimental temperature program:after the equilibration of the sample at −5° C., the samples were heatedthrough a modulated program from −5° C. to 210° C. at a rate of 1°C./min. On the basis of the classical recommendations, the set of MTDSCparameters chosen for analysing all specimens was amplitude of 0.5° C.and period of 80 seconds.

Experiments are performed using crimped aluminium sample pans or openpans in some cases. The quantity of product analysed is typicallybetween 2 and 13 mg.

Simultaneous TGA and DSC measurements are carried out using a SETSYSthermal analysis system produced by the Setaram corporation. A liquidnitrogen cooling system makes it possible to work at temperatures below20° C. The emitted gases, upon sample heating, are carried by a transfercapillary tube to a Pfeiffer mass spectrometer, the capillary tubeheated to 150° C. The mass spectrometer can analyze molecular fragmentsat <<m/e>> ratios between 1 and 200 uma. A sample with a mass between 1and 27 mg is contained in an open concave aluminum sample pan (75 μLcapacity) exposed to a nitrogen stream. The sample is heated at a rateof 1 or 5° C./min.

Water sorption measurements are performed on a DVS-1 automatedgravimetric vapor sorption analyser (Surface Measurement Systems Ltd.,London, UK). The DVS-1 measures the uptake and loss of vaporgravimetrically using a Cahn D200 recording ultra-microbalance with amass resolution of ±0.1 μg. The relative humidity around the sample iscontrolled by mixing saturated and dry carrier gas streams using massflow controllers. The temperature is maintained within ±0.1° C. byenclosing the entire system in a temperature-controlled incubator. Asample size between 15 and 20 mg is used. In general, the error in %(w/w) identified in a tested sample is ±0.1% (w/w).

Two protocols are utilized to operate the DVS. In one protocol, a sampleis dried at 0% relative humidity (RH) to remove any surface waterpresent and establish a dry, baseline mass. Next, the sample is exposedto the following relative humidity profile: 0%, 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 0%. In someinstances, this profile is repeated, yielding 4 sets of data going upand down in RH. In another protocol, the sample is first equilibrated at40% RH, then it was exposed to the following relative humidity profile:40%, 50%, 60%, 70%, 80%, 90%, 100%, 90%, 80%, 70%, 60%, 50% 40% 30%, 20%10%, 0%, 10%, 20%, 30%, 40%. At each stage, the sample mass is allowedto reach equilibrium before the relative humidity was increased ordecreased; equilibrium is met when the time rate of change of mass didnot exceed the value of 0.02%/min over a period of 30 or 60 minutes. Ifequilibrium state is not reached, the change in relative humidity tookplace automatically after 360 or 600 minutes. From the complete moisturesorption and desorption profile, an isotherm is calculated using the DVSAdvanced Analysis Suite v3.6. All experiments were performed at 25° C.

Some embodiments of the invention are described more specifically by theuse of the following examples, which are not intended in anyway to limitthe scope of the present invention.

EXAMPLES Example 1 Preparation of Different Polymorphs of the CitrateSalt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol

Eight different crystalline forms of the Citrate Salt are prepared bycrystallizing the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-olunder a variety conditions. In each instance, Form A of the Citrate Saltis exposed to a solvent or solvent system under the particularconditions listed in Table 9 to generate the particular form(s) oramorphous Citrate Salt.

TABLE 9 Crystallization Conditions for Various Samples of the CitrateSalt Form(s) identified Crystallization Conditions B Crystallizationafter dissolution at high temperature in ethanol. B Crystallizationafter dissolution at ambient temperature in methanol/heptane. BCrystallization after dissolution in MEK/water and azeotropicdistillation (isolation in MEK). B Crystallization after dissolution inn-propanol, followed by a slow evaporation at ambient temperature. A + CCrystallization after dissolution at high temperature in THF/water. AMaturation in toluene at ambient temperature. B Crystallization afterdissolution at high temperature in isopropanol. B Crystallization afterdissolution at high temperature in isopropanol/water. B Crystallizationafter dissolution at high temperature in n-propanol. A + DCrystallization after dissolution THF, followed by a slow evaporation atambient temperature and atmospheric pressure. B Crystallization afterdissolution in azeotropic mixture acetonitrile/water. AmorphousDissolution at ambient temperature in DMSO/water. Amorphous Dissolutionin dichloromethane/methanol, followed by a quick evaporation at ambienttemperature and low pressure. Amorphous Dissolution in THF/water andazeotropic distillation (isolation in water). E + F + G Maturation inwater of the amorphous form at ambient temperature. E Maturation inwater of Form A at ambient temperature. B Maturation in ethanol of FormsA and B at ambient temperature. A + C Maturation in THF of Forms A and Bat ambient temperature. B Maturation in methanol of Forms A and B atambient temperature.

Example 2 Preparation of Form A of the Citrate Salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol

The tartrate of (S)-4-(4-Chloro-phenyl)-3,3-dimethylpiperidin-4-ol (7.5kg) is mixed with2-[5-(3-Bromo-propylidene)-5,11-dihydro-10-oxa-1-aza-dibenzo[a,d]cyclohepten-7-yl]-propan-2-ol(7.5 kg) and acetonitrile (90 liters) in a 250 liter glass-lined reactorinerted with nitrogen gas. The (S)-isomer is obtained by mixing4-(4-Chloro-phenyl)-3,3-dimethylpiperidin-4-ol with an excess of(R,S)-tartaric acid to precipitate the (S) tartrate. The mixture isagitated with an impeller and cooled to between 0° C. and 5° C.

Water (23 liters) is charged to the vessel, followed by potassiumcarbonate (9.56). The solution is warmed to 20±5° C.; the temperaturewas maintained for at least 3 days. HPLC is utilized to follow thecourse of the reaction.

Water (45 liter) is added to the reactor. The total contents areconcentrated using vacuum distillation at a pressure selected tomaintain the temperature below 40° C. When the total contents volume isabout 60 liters, the distillation is halted. The contents are brought toatmospheric pressure and a temperature between 20° C. to 30° C.

Next, dichloromethane 62 liters is charged to the reactor; the reactoris agitated for 15 minutes. The total volume is then allowed to sit for15 minutes. The aqueous phase is subsequently discarded, and the organicphase returned to the reactor. The organic phase is twice washed withwater (45 liters each time); each wash including agitating for 15minutes, sitting for 15 minutes, and discarding the upper aqueous phase.

Di-tert-butyl dicarbonate (0.12 kg.) in dichloromethane (4 liters) isadded to the reactor. The doseur is rinsed with 4 liters ofdichloromethane. The solution is maintained at 20° C. for 10-20 minutes.The solution is analyzed for the presence of the tartrate of(S)-4-(4-Chloro-phenyl)-3,3-dimethylpiperidin-4-ol. If the tartrate isstill present, di-tert-butyl dicarbonate was added in a 2:1 ratio molarratio with the remaining tartrate. In this particular instance, moredi-tert butyl dicarbonate (0.12 kg) in dichloromethane (4 liter) wasrequired.

The contents of the reactor are again concentrated by vacuumdistillation, maintaining the temperature at or below 40° C., until theresidual volume is about 40 liters. The remaining contents are broughtto a temperature between 20° C. and 30° C., and atmospheric pressure.

Next, acetone (31 liters) is charged to the reactor. The reactor'scontents are concentrated to 40 liters by vacuum distillation whilemaintaining the temperature at or below 40° C. The remaining volume isthen brought to a temperature between 20° C. and 30° C., and atmosphericpressure. The solvent exchange with acetone is repeated three moretimes.

Next, acetone (58 liters) and an acetonic slurry of decolorizing carbon(L3S, 0.38 kg of carbon; 2 liters acetone) is added to the reactor. Themixture is held for one hour at 20±5° C. The solution is filteredthrough a 0.22 um solvex-type filter, the filtrate being sent to a 160liter reactor.

Citric acid (3.68 kg) and acetone (24 liters) are charged to the 250liter reactor. The mixture is agitated at 20±5° C. until a solution wasformed. The contents are heated to 35° C., and then transferred to the160 liter reactor. The temperature of the 160 liter reactor ismaintained at 35±5° C. during the transfer. The product begins tocrystallize during the transfer. The temperature of 35±5° C. ismaintained for 10 to 20 minutes after the transfer is complete.

The crystals and solution dispersion is filtered (filter drier; cake:d=55 cm, h=13 cm; filter time 4 hours). The filter cake is twice washedwith acetone (34 liters of acetone each wash; wash time ˜10 hourstotal). The product is dried in a filter drier (40±5° C./−0.99 bar,non-agitated, ca. 100 hours) to produce 10.42 kg product (75% yield),which assays to 99% (w/w) of the citrate salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.

Example 3 Preparation of Form B of the Citrate Salt of(S)-4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol

The tartrate of (S)-4-(4-Chloro-phenyl)-3,3-dimethylpiperidin-4-ol (20kg) is mixed with2-[5-(3-Bromo-propylidene)-5,11-dihydro-10-oxa-1-aza-dibenzo[a,d]cyclohepten-7-yl]-propan-2-ol(20 kg) and acetonitrile (240 liters) in a 600 liter glass-lined reactorinerted with nitrogen gas. The mixture is agitated with an impeller andcooled to between 0° C. and 5° C.

Water (60 liters) is charged to the vessel, followed by addition ofpotassium carbonate 25.5 kg. The solution is warmed to 20±5° C.; thetemperature was maintained for about 4 days. HPLC is utilized to followthe course of the reaction.

Water (120 liters) is added to the reactor. The total contents areconcentrated using vacuum distillation at a pressure selected tomaintain the temperature below 40° C. When the total volume is about 90liters, the distillation is halted. The remaining volume is brought to atemperature between 20° C. and 30° C. and atmospheric pressure.

Next, dichloromethane (165 liters) is charged to the reactor; thereactor is then agitated for 15 minutes. The total volume issubsequently allowed to sit for 15 minutes. The aqueous phase isdiscarded, and the organic phase returned to the reactor. The organicphase is twice washed with water (120 liters each time); each washincluding agitating for 15 minutes, sitting for 15 minutes, anddiscarding the upper aqueous phase.

The organic phase is analyzed for the presence of the tartrate of(S)-4-(4-Chloro-phenyl)-3,3-dimethylpiperidin-4-ol. Di-tert-butyldicarbonate in dichloromethane is added to the reactor in an approximateamount of 2 moles of dicarbonate for each mole of tartrate remaining. Inthis particular instance, about 0.3 kg of dicarbonate is used with about5 liters of dichloromethane. The doseur is rinsed with 5 liters ofdichloromethane. The solution is maintained at 20° C. for 10 to 20minutes. The solution is again analyzed for the presence of thetartrate. If the tartrate is still present, more di-tert-butyldicarbonate in dichloromethane is added to the reactor in an amount ofdicarbonate to tartrate of 2:1 on a molar basis.

The contents of the reactor are again concentrated by vacuumdistillation, maintaining the temperature at or below 40° C., until theresidual volume is about 90 liters. The remaining contents are broughtto a temperature between 20° C. and 30° C., and atmospheric pressure.

Next, absolute ethanol (71 liters) is charged to the reactor. Thereactor's contents are concentrated to 90 liters by vacuum distillationwhile maintaining the temperature at or below 40° C. The remainingvolume is then brought to a temperature 20° C. and 30° C. between andatmospheric pressure. The solvent exchange with absolute ethanol isrepeated two more times.

Next, ethanol (71 liters) and an ethanolic slurry of decolorizing carbon(L3S, 1.15 kg of carbon; ethanol, qsp) are added to the reactor. Themixture is held for one hour at 20±5° C. The solution is filteredthrough a 0.22 um solvex-type filter, the filtrate being sent to a 400liter reactor.

Citric acid (9.85 kg) and ethanol (37 liters) are charged to a 100 literhastelloy reactor. The mixture is agitated at 20±5° C. until a solutionis formed, the citric acid solution being transferred to a drumsubsequently. The citric acid solution (13 kg) is added to the 400 literreactor containing the filtrate, while maintaining a temperature of20±2° C. in the reactor. The reactor is seeded with a slurry of Form Bcrystals of the Citrate Salt made from a portion of the solution in the400 liter reactor. The crystals are milled for 30 seconds in aThurrax-type mixer in liquid from the 400 liter reactor. The seededcrystallization mixture is maintained for 30 minutes.

The remainder of the citric acid solution is transferred to the 400liter reactor over a 2.5 to 3 hour time period, while maintaining thetemperature in the reactor at 20±2° C. The product crystallized duringthis transfer. Any excess citric acid solution is rinsed into thereactor with 5 liters of ethanol.

The reactor is cooled to 0±2° C. over 30 minutes; this temperature issubsequently maintained for at least 1 hour. The suspension issubsequently filtered (Nutsch filter, filter time 1 hour) to produce afilter cake. The cake is twice rinsed with ethanol (55 liters eachrinse; wash time ˜36 hours total). The cake is dried in a tray drier(40±5° C./−0.99 bar, non-agitated, ca. 48 hours) to produce 27.3 kg. ofproduct (73% yield), which assays to 97% (w/w) of the citrate salt of4-(4-Chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A citrate salt of(S)-4-(4-chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,having structural formula (III):

wherein the citrate salt is crystalline Form A.
 2. A citrate salt of(S)-4-(4-chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,having structural formula (III):

wherein the citrate salt is crystalline Form B.
 3. The citrate salt ofclaim 1, wherein the single crystalline form is characterized by atleast one of the x-ray powder diffraction peaks at 2θ angles of 9.8,11.7, 12.6, 15.5, 15.7, 15.9, 17.3, 17.5, 18.2, 19.0, and 19.7.
 4. Thecitrate salt of claim 1, wherein the single crystalline form ischaracterized by a x-ray powder diffraction pattern substantiallysimilar to FIG.
 1. 5. The citrate salt of claim 2, wherein the singlecrystalline form is characterized by at least one of the x-ray powderdiffraction peaks at 2θ angles of 10.6, 11.6, 12.3, 14.8, 15.8, 16.1,16.7, 18.8, 20.6, 21.7, and 24.5.
 6. The citrate salt of claim 2,wherein the single crystalline form is characterized by a x-ray powderdiffraction pattern substantially similar to FIG.
 6. 7. A pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or diluent;and a citrate salt of(S)-4-(4-chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,wherein the citrate salt is crystalline Form A.
 8. A pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or diluent;and a citrate salt of(S)-4-(4-chloro-phenyl)-1-{3-[7-(1-hydroxy-1-methyl-ethyl)-11H-10-oxa-1-aza-dibenzo[a,d]cyclohepten-5-ylidene]-propyl}-3,3-dimethyl-piperidin-4-ol,wherein the citrate salt is crystalline Form B.
 9. The pharmaceuticalcomposition of claim 7, wherein the single crystalline form ischaracterized by at least one of the x-ray powder diffraction peaks at2θ angles of 9.8, 11.7, 12.6, 15.5, 15.7, 15.9, 17.3, 17.5, 18.2, 19.0,and 19.7.
 10. The pharmaceutical composition of claim 7, wherein thesingle crystalline form is characterized by a x-ray powder diffractionpattern substantially similar to FIG.
 1. 11. The pharmaceuticalcomposition of claim 8, wherein the single crystalline form ischaracterized by at least one of the x-ray powder diffraction peaks at2θ angles of 10.6, 11.6, 12.3, 14.8, 15.8, 16.1, 16.7, 18.8, 20.6, 21.7,and 24.5.
 12. The pharmaceutical composition of claim 8, wherein thesingle crystalline form is characterized by a x-ray powder diffractionpattern substantially similar to FIG.
 6. 13. The citrate salt of claim1, wherein the citrate salt is at least 90% crystalline Form A.
 14. Thecitrate salt of claim 2, wherein the citrate salt is at least 90%crystalline Form B.
 15. The pharmaceutical composition of claim 7,wherein the citrate salt is at least 90% crystalline Form A.
 16. Thepharmaceutical composition of claim 8, wherein the citrate salt is atleast 90% crystalline Form B.